Biomarkers for the diagnosis and treatment of fibrotic lung disease

ABSTRACT

The present disclosure provides a method of treating a fibrotic lung disease in a subject comprising administering to the subject an effective amount of a therapeutic agent, wherein the subject is asymptomatic and wherein the subject is at risk of developing the fibrotic lung disease.

RELATED APPLICATIONS

This application claims the benefit of provisional application U.S. Ser. No. 62/525,087, filed Jun. 26, 2017, and U.S. Ser. No. 62/525,088, filed Jun. 26, 2017, the contents of each of which are herein incorporated by reference in their entirety.

INCORPORATION OF SEQUENCE LISTING

The contents of the text file named “UNCO-018_001WO_SeqList_ST25.txt,” which was created on Jun. 21, 2018 and is 418 KB in size, are hereby incorporated by reference in their entirety.

FIELD OF THE DISCLOSURE

The disclosure is directed to molecular biology, genetics, and therapeutics for fibrotic lung disease.

BACKGROUND

Fibrotic pulmonary diseases are progressive and irreversible. Standard therapies are mere palliative as they cannot address the underlying disease mechanism once the subject has progressed to a point at which symptoms are present. Thus, there is a long-felt but unmet need in the field for a method of treating asymptomatic subjects as well as those who are at risk of developing fibrotic pulmonary diseases to prevent onset of the disease, delay onset of the disease, or reduce the severity of disease symptoms, The methods of the disclosure provide a preventative or efficacious treatment, as opposed to a merely palliative treatment, for asymptomatic subjects as well as those subjects at risk of developing the disease.

SUMMARY

The disclosure provides a method of treating a fibrotic lung disease in a subject comprising administering to the subject an effective amount of a therapeutic agent, wherein the subject is asymptomatic and wherein the subject is at risk of developing the fibrotic lung disease.

In some embodiments of the methods of the disclosure, the subject presents radiographic Usual Interstitial Pneumonia (UIP). In some embodiments, the subject has fibrotic interstitial lung disease (FILD). In some embodiments, the subject has a blood relative with familial interstitial pneumonia (FIP). In some embodiments, including those embodiments wherein the subject has a blood relative with familial interstitial pneumonia (FIP), the blood relative is a sibling. Alternatively, or in addition, in some embodiments, the subject has a mutation in a sequence encoding Mucin 5B (MUC5B), Telomerase RNA Component (TERC), Family with sequence similarity 13 member A (FAM13A), Telomerase Reverse Transcriptase (TERT), Desmoplakin (DSP), Zinc-alpha 2-Glycoprotein 1 (AZGP1), Oligonucleotide/oligosaccharide-binding Fold Containing 1 (OBFC1), ATPase Phospholipid Transporting 11A (ATP11A), Isovaleryl-CoA dehydrogenase (IVD)/Dispatched RND Transporter Family Member 2 (DISP2), Dipeptidyl Peptidase 9 (DPP9), Sialic Acid Binding Ig-Like Lectin 14 (SIGLEC14), Adrenomedullin 2 (ADM2), Tetraspanin 5 (TSPAN5), Calcium/Calmodulin-Dependent Protein Kinase 1 (CAMKK1), zinc figner with KRAB and SCAN domains 1 (ZKSCAN1), isovaleryl-CoA dehydrogenase (IVD), ATPase phospholipid transporting 11A (AK025511) or Matrix Metalloprotease-7 (MMP-7).

In some embodiments of the methods of the disclosure, the subject has a mutation in a sequence encoding MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7.

In some embodiments of the methods of the disclosure, the subject has a mutation in a nucleic acid or amino acid sequence encoding a gene or gene product that is upregulated in a subject having a fibrotic pulmonary disease of the disclosure. In some embodiments of the methods of the disclosure, the subject has a mutation in a nucleic acid or amino acid sequence encoding Leukotriene A4 Hydrolase (LTA4H), Surfactant Protein B (SFTPB), Breast Cancer Anti-Estrogen Resistance 3 (BCAR3), C-X-C motif Chemokine Ligand 13 (CXCL13), EPH Receptor A2 (EPHA2), Serum Amyloid A1 (SAA1), Phospholipase A2 Group IIA (PLA2G2A), Insulin-Like Growth Factor Binding Protein 3 (IGFBP3), C-C Motif Chemokine Ligand 28 (CCL28), S100 Calcium Binding Protein A12 (S100A12), Thromboxane A Synthase 1 (TBXAS1), Leukocyte Cell Derived Chemotaxin 1 (LECT1), Complement C3 (C3), Gastrin Releasing Peptide (GRP), C-Reactive Protein (CRP), Vitrin (VIT), Insulin-Like Growth Factor Binding Protein 1 (IGFBP1), Family with Sequence Similarity 173 Member A (FAM173A), Natriuretic Peptide A (NPPA), Secreted Frizzled Related Protein 1 (SFRP1), Ezrin (EZR), Inter-Alpha-Trypsin Inhibitor Heavy Chain Family Member 5 (ITIH5), Pleckstrin and Sec7 Domain Containing 2 (PSD2), Galectin 3 Binding Protein (LGALS3BP), Catenin Beta 1 (CTNNB1), Chromodomain Y Like 2 (CDYL2), Matrix Metallopeptidase 7 (MMPI), Apolipoprotein B (APOB), Proline and Arginine Rich End Leucine Rich Repeat Protein (PRELP), Eukaryotic Translation Initiation Factor 1A, X-linked (EIF1AX), Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF), TNF Receptor Superfamily Member 13C (TNFRSF13C), Deformed Epidermal Autoregulatory Factor 1 transcription factor (DEAF1), Tumor Protein Translationally-Controlled 1 (TPT1), Unc-5 Netrin Receptor B (UNC5B), Phosphatidylethanolamine Binding Protein 1 (PEBP1), Syntaxin 8 (STX8), Polymeric Immunoglobulin Receptor (PIGR), Adenine Phosphoribosyltransferase (APRT), Matrix Metallopeptidase 3 (MMP3), Galectin 7 (LGALS7), Bruton Tyrosine Kinase (BTK), NSFL1 Cofactor (NSFL1C), FER Tyrosine Kinase (FER), Regenerating Family Member 1 Beta (REG1B), SMAD Family Member 2 (SMAD2), Interleukin 1 Receptor Like 1 (IL1RL1), C-C Motif Chemokine Ligand 18 (CCL18), Acid Phosphatase 2 Lysosomal (ACP2), Eukaryotic Translation Initiation Factor 4E Family Member 2 (EIF4E2), Neurexin 3 (NRXN3), IGF Like Family Member 1 (IGFL1), NME/NM23 Nucleoside Diphosphate Kinase 1 (NME1), Potassium Voltage-Gated Channel Isk-Related Family Member 1-Like (KCNE1L) or Neurexophilin 2 (NXPH2).

In some embodiments of the methods of the disclosure, the subject has a mutation in a nucleic acid or amino acid sequence encoding a gene or gene product that is downregulated in a subject having a fibrotic pulmonary disease of the disclosure. In some embodiments of the methods of the disclosure, the subject has a mutation in a nucleic acid or amino acid sequence encoding Surfactant Protein D (SFTPD), Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), Histone Cluster 1 H1 Family Member C (HIST1H1C), YTH Domain Containing 1 (YTHDC1), Plexin A1 (PLXNA1), Serine Peptidase Inhibitor Kazal Type 6 (SPINK6), LDL Receptor Related Protein Associated Protein 1 (LRPAP1), Secretoglobin Family 3A Member 1 (SCGB3A1), H2A Histone Family Member Z (H2AFZ) or Chromosome 1 Open Reading Frame 162 (Clorf162).

In some embodiments of the methods of the disclosure, the subject has a mutation in a sequence encoding MUC5B. In some embodiments, the mutation is a polymorphism in a sequence encoding a MUC5B promoter. In some embodiments, the polymorphism is rs35705950 comprising (SEQ ID NO: 7).

In some embodiments of the methods of the disclosure, the human subject has a mutation in a sequence encoding TERC. In some embodiments, the mutation is a polymorphism in a sequence encoding TERC or a regulatory sequence thereof. In some embodiments the polymorphism is rs6793295 comprising (SEQ ID NO: 1).

In some embodiments of the methods of the disclosure, the human subject has a mutation in a sequence encoding intronic FAM13A. In some embodiments, the mutation is a polymorphism in a sequence encoding intronic FAM13A or a regulatory sequence thereof. In some embodiments, the polymorphism is rs2609260.

In some embodiments of the methods of the disclosure, the human subject has a mutation in a sequence encoding intronic TERT. In some embodiments, the mutation is a polymorphism in a sequence encoding intronic TERT or a regulatory sequence thereof. In some embodiments, the polymorphism is rs4449583.

In some embodiments of the methods of the disclosure, the human subject has a mutation in a sequence encoding intronic DSP. In some embodiments, the mutation is a polymorphism in a sequence encoding intronic DSP or a regulatory sequence thereof. In some embodiments, the polymorphism is rs2076295.

In some embodiments of the methods of the disclosure, the human subject has a mutation in a sequence encoding intronic ZKSCAN1. In some embodiments, the mutation is a polymorphism in a sequence encoding intronic ZKSCAN1 or a regulatory sequence thereof. In some embodiments, the polymorphism is rs6963345.

In some embodiments of the methods of the disclosure, the human subject has a mutation in a sequence encoding intronic OBFC1. In some embodiments, the mutation is a polymorphism in a sequence encoding intronic OBFC1 or a regulatory sequence thereof. In some embodiments, the polymorphism is rs2488000.

In some embodiments of the methods of the disclosure, the human subject has a mutation in a sequence encoding an AK025511 3′ UTR. In some embodiments, the mutation is a polymorphism in a sequence encoding an AK025511 3′ UTR or a regulatory sequence thereof. In some embodiments, the polymorphism is rs1278769.

In some embodiments of the methods of the disclosure, the human subject has a mutation in a sequence encoding IVD. In some embodiments, the mutation is a polymorphism in a sequence encoding intronic IVD or a regulatory sequence thereof. In some embodiments, the polymorphism is rs35700143.

In some embodiments of the methods of the disclosure, the human subject has a mutation in a sequence encoding intronic DPP9. In some embodiments, the mutation is a polymorphism in a sequence encoding intronic DPP9 or a regulatory sequence thereof. In some embodiments, the polymorphism is rs12610495.

In some embodiments of the methods of the disclosure, the subject has a mutation in a sequence encoding FAM13A. In some embodiments, the mutation is a polymorphism in a sequence encoding FAM13A or a regulatory sequence thereof. In some embodiments the polymorphism is rs2609255 comprising (SEQ ID NO: 2).

In some embodiments of the methods of the disclosure, the subject has a mutation in a sequence encoding TERT. In some embodiments, the mutation is a polymorphism in a sequence encoding TERT or a regulatory sequence thereof. In some embodiments the polymorphism is rs2736100 comprising (SEQ ID NO: 3).

In some embodiments of the methods of the disclosure, the subject has a mutation in a sequence encoding DSP. In some embodiments, the mutation is a polymorphism in a sequence encoding DSP or a regulatory sequence thereof. In some embodiments the polymorphism is rs2076295 comprising (SEQ ID NO: 4).

In some embodiments of the methods of the disclosure, the subject has a mutation in a sequence encoding AZGP1. In some embodiments, the mutation is a polymorphism in a sequence encoding AZGP1 or a regulatory sequence thereof. In some embodiments the polymorphism is rs4727443 comprising (SEQ ID NO: 5).

In some embodiments of the methods of the disclosure, the subject has a mutation in a sequence encoding OBFC1. In some embodiments, the mutation is a polymorphism in a sequence encoding OBFC1 or a regulatory sequence thereof. In some embodiments the polymorphism is rs11191865 comprising (SEQ ID NO: 6).

In some embodiments of the methods of the disclosure, the subject has a mutation in a sequence encoding ATP11A. In some embodiments, the mutation is a polymorphism in a sequence encoding ATP11A or a regulatory sequence thereof. In some embodiments the polymorphism is rs12787690 comprising (SEQ ID NO: 8).

In some embodiments of the methods of the disclosure, the subject has a mutation in a sequence encoding IVD/DISP2. In some embodiments, the mutation is a polymorphism in a sequence encoding IVD/DISP2 or a regulatory sequence thereof. In some embodiments the polymorphism is rs2034650 comprising (SEQ ID NO: 9).

In some embodiments of the methods of the disclosure, the subject has a mutation in a sequence encoding DPP9. In some embodiments, the mutation is a polymorphism in a sequence encoding DPP9 or a regulatory sequence thereof. In some embodiments the polymorphism is rs12610495 comprising (SEQ ID NO: 10).

In some embodiments of the methods of the disclosure, the fibrotic lung disease is pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), an interstitial lung abnormality (ILA), or an asymptomatic ILA. In some embodiments, the fibrotic lung disease is pulmonary fibrosis or IPF. In some embodiments, the fibrotic lung disease is IPF.

In some embodiments of the methods of the disclosure, the therapeutic agent comprises a N-acetylcysteine, pirfenidone, and nintedanib.

In some embodiments of the methods of the disclosure, the therapeutic agent comprises pirfenidone. In some embodiments, the effective dosage is administered orally as a capsule or a tablet. In some embodiments, including those embodiments wherein the therapeutic agent comprises pirfenidone, the effective dosage is about 2400 mg/day. In some embodiments, the effective dosage is administered according to an escalating dosage regimen. In some embodiments, including those embodiments wherein the therapeutic agent comprises pirfenidone, the escalating dosage regimen comprises (a) administering to the subject about 800 mg of pirfenidone per day for a first week; (b) administering to the subject about 1600 mg of pirfenidone per day for a second week; and (c) administering to the subject about 2400 mg of pirfenidone per day for the remainder of the treatment. In some embodiments, including those embodiments wherein the therapeutic agent comprises pirfenidone, the escalating dosage regimen comprises (a) administering to the subject a capsule or tablet comprising about 250 mg of pirfenidone three times a day for a first week; (b) administering to the subject two capsules or tablets comprising about 250 mg of pirfenidone three times a day for a second week; and (c) administering to the subject three capsules or tablets comprising about 250 mg of pirfenidone three times a day for the remainder of the treatment. In some embodiments of the escalating dosage regimen, the capsule or tablet comprises 267 mg of pirfenidone.

In some embodiments of the methods of the disclosure, the therapeutic agent comprises nintedanib. In some embodiments, the effective dosage is administered orally as a capsule or a tablet. In some embodiments, including those embodiments wherein the therapeutic agent comprises nintedanib, the effective dosage is about 300 mg/day. In some embodiments, the effective dosage is about 150 mg administered twice per day, wherein the daily doses are administered about 12 hours apart from one another. In some embodiments, including those embodiments wherein the therapeutic agent comprises nintedanib, the effective dosage is about 200 mg/day. In some embodiments, the effective dosage is about 100 mg administered twice per day, wherein the daily doses are administered about 12 hours apart from one another. In some embodiments, including those embodiments wherein the therapeutic agent comprises nintedanib, the effective dosage is administered according to a modified or interrupted dosage regimen. In some embodiments, the modified or interrupted dosage regimen comprises (a) administering to the subject about 300 mg of nintedanib per day until the subject presents an elevated level of liver enzymes compared to a control level of liver enzymes; (b) administering to the subject about 200 mg of nintedanib per day until the subject presents the control level of liver enzymes; and (c) administering to the subject about 300 mg of nintedanib per day for the remainder of the treatment; wherein the control level of liver enzymes is a level detected in the subject prior to an initiation of the treatment. In some embodiments, including those embodiments wherein the therapeutic agent comprises nintedanib, the modified or interrupted regimen comprises (a) administering to the subject a capsule or tablet comprising about 150 mg of nintedanib twice per day until the subject presents an elevated level of liver enzymes compared to a control level of liver enzymes; (b) administering to the subject two capsules or tablets comprising about 100 mg twice per day until the subject presents an elevated level of liver enzymes compared to a control level of liver enzymes; and (c) administering to the subject a capsule or tablet comprising about 150 mg of nintedanib twice per day for the remainder of the treatment; wherein the control level of liver enzymes is a level detected in the subject prior to an initiation of the treatment.

In some embodiments of the methods of the disclosure, the therapeutic agent prevents the onset or development of a sign or symptom of the fibrotic lung disease.

In some embodiments of the methods of the disclosure, the therapeutic agent delays the onset or development of a sign or symptom of the fibrotic lung disease when compared to the expected onset of the sign or symptom in the absence of treatment with the therapeutic agent.

In some embodiments of the methods of the disclosure, the therapeutic agent reduces the severity of a sign or symptom of the fibrotic lung disease when compared to the expected severity of the sign or symptom in the absence of treatment with the therapeutic agent.

In some embodiments of the methods of the disclosure, the therapeutic agent reduces the severity of a sign or symptom of the fibrotic lung disease when compared to the expected severity of the sign or symptom in the absence of treatment with the therapeutic agent.

In some embodiments of the methods of the disclosure, the at least one sign of the fibrotic lung disease is detectable before the subject presents a symptom of the fibrotic lung disease. In some embodiments, the at least one sign comprises gradual or unintended weight loss, clubbing of the fingers or toes, rapid and shallow breathing, fibrotic lesions in one or both lungs detectable by radiography, or a cough. In some embodiments, the symptom comprises shortness of breath during exercise, shortness of breath at rest, a dry and hacking cough, repeated bouts of coughing, and uncontrollable bouts of coughing.

In some embodiments of the methods of the disclosure, the method prevents the onset of a secondary condition associated with a severe form of the fibrotic lung disease. In some embodiments, a secondary condition comprises a collapsed lung, an infected lung, a blood clot in a lung, lung cancer, respiratory failure, pulmonary hypertension, heart failure or death.

The disclosure provides a method of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease, comprising administering to a non-human subject a dose of a composition that modifies transcription or translation of a sequence encoding Mucin 5B (MUC5B), Telomerase RNA Component (TERC), Family with sequence similarity 13 member A (FAM13A), Telomerase Reverse Transcriptase (TERT), Desmoplakin (DSP), Zinc-alpha 2-Glycoprotein 1 (AZGP1), Oligonucleotide/oligosaccharide-binding Fold Containing 1 (OBFC1), ATPase Phospholipid Transporting 11A (ATP11A), Isovaleryl-CoA dehydrogenase (IVD)/Dispatched RND Transporter Family Member 2 (DISP2), Dipeptidyl Peptidase 9 (DPP9), Sialic Acid Binding Ig-Like Lectin 14 (SIGLEC14), Adrenomedullin 2 (ADM2), Tetraspanin 5 (TSPAN5), Calcium/Calmodulin-Dependent Protein Kinase Kinase 1 (CAMKK1) or Matrix Metalloprotease-7 (MMP-7), wherein the dose of the composition is tolerable to the non-human subject and wherein the dose of the composition is therapeutically effective.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the method of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease, comprising administering to a non-human subject a composition that modifies an activity of a product of a sequence encoding MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7, wherein the dose of the composition is tolerable to the non-human subject and wherein the dose of the composition is therapeutically effective.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the composition that modifies transcription or translation decreases or inhibits transcription or translation.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the composition decreases or inhibits transcription or translation of a sequence encoding a gene selected from the group consisting of Leukotriene A4 Hydrolase (LTA4H), Surfactant Protein B (SFTPB), Breast Cancer Anti-Estrogen Resistance 3 (BCAR3), C—X—C motif Chemokine Ligand 13 (CXCL13), EPH Receptor A2 (EPHA2), Serum Amyloid A1 (SAA1), Phospholipase A2 Group IIA (PLA2G2A), Insulin-Like Growth Factor Binding Protein 3 (IGFBP3), C-C Motif Chemokine Ligand 28 (CCL28), S100 Calcium Binding Protein A12 (S100A12), Thromboxane A Synthase 1 (TBXAS1), Leukocyte Cell Derived Chemotaxin 1 (LECT1), Complement C3 (C3), Gastrin Releasing Peptide (GRP), C-Reactive Protein (CRP), Vitrin (VIT), Insulin-Like Growth Factor Binding Protein 1 (IGFBP1), Family with Sequence Similarity 173 Member A (FAM173A), Natriuretic Peptide A (NPPA), Secreted Frizzled Related Protein 1 (SFRP1), Ezrin (EZR), Inter-Alpha-Trypsin Inhibitor Heavy Chain Family Member 5 (ITIH5), Pleckstrin and Sec7 Domain Containing 2 (PSD2), Galectin 3 Binding Protein (LGALS3BP), Catenin Beta 1 (CTNNB1), Chromodomain Y Like 2 (CDYL2), Matrix Metallopeptidase 7 (MMPI), Apolipoprotein B (APOB), Proline and Arginine Rich End Leucine Rich Repeat Protein (PRELP), Eukaryotic Translation Initiation Factor 1A, X-linked (EIF1AX), Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF), TNF Receptor Superfamily Member 13C (TNFRSF13C), Deformed Epidermal Autoregulatory Factor 1 transcription factor (DEAF1), Tumor Protein Translationally-Controlled 1 (TPT1), Unc-5 Netrin Receptor B (UNCSB), Phosphatidylethanolamine Binding Protein 1 (PEBP1), Syntaxin 8 (STX8), Polymeric Immunoglobulin Receptor (PIGR), Adenine Phosphoribosyltransferase (APRT), Matrix Metallopeptidase 3 (MMP3), Galectin 7 (LGALS7), Bruton Tyrosine Kinase (BTK), NSFL1 Cofactor (NSFL1C), FER Tyrosine Kinase (FER), Regenerating Family Member 1 Beta (REG1B), SMAD Family Member 2 (SMAD2), Interleukin 1 Receptor Like 1 (IL1RL1), C-C Motif Chemokine Ligand 18 (CCL18), Acid Phosphatase 2 Lysosomal (ACP2), Eukaryotic Translation Initiation Factor 4E Family Member 2 (EIF4E2), Neurexin 3 (NRXN3), IGF Like Family Member 1 (IGFL1), NME/NM23 Nucleoside Diphosphate Kinase 1 (NME1), Potassium Voltage-Gated Channel Isk-Related Family Member 1-Like (KCNE1L) or Neurexophilin 2 (NXPH2).

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the composition that modifies transcription or translation increases or activates transcription or translation.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the composition increases or activates transcription or translation of a sequence encoding a gene selected from the group consisting of Surfactant Protein D (SFTPD), Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), Histone Cluster 1 H1 Family Member C (HIST1H1C), YTH Domain Containing 1 (YTHDC1), Plexin A1 (PLXNA1), Serine Peptidase Inhibitor Kazal Type 6 (SPINK6), LDL Receptor Related Protein Associated Protein 1 (LRPAP1), Secretoglobin Family 3A Member 1 (SCGB3A1), H2A Histone Family Member Z (H2AFZ) or Chromosome 1 Open Reading Frame 162 (Clorf162).

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the composition that modifies an activity decreases or inhibits the activity.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the composition decreases or inhibits the activity of a sequence encoding a gene selected from Leukotriene A4 Hydrolase (LTA4H), Surfactant Protein B (SFTPB), Breast Cancer Anti-Estrogen Resistance 3 (BCAR3), C—X—C motif Chemokine Ligand 13 (CXCL13), EPH Receptor A2 (EPHA2), Serum Amyloid A1 (SAA1), Phospholipase A2 Group IIA (PLA2G2A), Insulin-Like Growth Factor Binding Protein 3 (IGFBP3), C-C Motif Chemokine Ligand 28 (CCL28), S100 Calcium Binding Protein A12 (S100A12), Thromboxane A Synthase 1 (TBXAS1), Leukocyte Cell Derived Chemotaxin 1 (LECT1), Complement C3 (C3), Gastrin Releasing Peptide (GRP), C-Reactive Protein (CRP), Vitrin (VIT), Insulin-Like Growth Factor Binding Protein 1 (IGFBP1), Family with Sequence Similarity 173 Member A (FAM173A), Natriuretic Peptide A (NPPA), Secreted Frizzled Related Protein 1 (SFRP1), Ezrin (EZR), Inter-Alpha-Trypsin Inhibitor Heavy Chain Family Member 5 (ITIH5), Pleckstrin and Sec7 Domain Containing 2 (PSD2), Galectin 3 Binding Protein (LGALS3BP), Catenin Beta 1 (CTNNB1), Chromodomain Y Like 2 (CDYL2), Matrix Metallopeptidase 7 (MMPI), Apolipoprotein B (APOB), Proline and Arginine Rich End Leucine Rich Repeat Protein (PRELP), Eukaryotic Translation Initiation Factor 1A, X-linked (EIF1AX), Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF), TNF Receptor Superfamily Member 13C (TNFRSF13C), Deformed Epidermal Autoregulatory Factor 1 transcription factor (DEAF1), Tumor Protein Translationally-Controlled 1 (TPT1), Unc-5 Netrin Receptor B (UNC5B), Phosphatidylethanolamine Binding Protein 1 (PEBP1), Syntaxin 8 (STX8), Polymeric Immunoglobulin Receptor (PIGR), Adenine Phosphoribosyltransferase (APRT), Matrix Metallopeptidase 3 (MMP3), Galectin 7 (LGALS7), Bruton Tyrosine Kinase (BTK), NSFL1 Cofactor (NSFL1C), FER Tyrosine Kinase (FER), Regenerating Family Member 1 Beta (REG1B), SMAD Family Member 2 (SMAD2), Interleukin 1 Receptor Like 1 (IL1RL1), C-C Motif Chemokine Ligand 18 (CCL18), Acid Phosphatase 2 Lysosomal (ACP2), Eukaryotic Translation Initiation Factor 4E Family Member 2 (EIF4E2), Neurexin 3 (NRXN3), IGF Like Family Member 1 (IGFL1), NME/NM23 Nucleoside Diphosphate Kinase 1 (NME1), Potassium Voltage-Gated Channel Isk-Related Family Member 1-Like (KCNE1L) or Neurexophilin 2 (NXPH2).

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the composition that modifies an activity increases or activates the activity.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the composition increases or activates the activity of a sequence encoding Surfactant Protein D (SFTPD), Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), Histone Cluster 1 H1 Family Member C (HIST1H1C), YTH Domain Containing 1 (YTHDC1), Plexin A1 (PLXNA1), Serine Peptidase Inhibitor Kazal Type 6 (SPINK6), LDL Receptor Related Protein Associated Protein 1 (LRPAP1), Secretoglobin Family 3A Member 1 (SCGB3A1), H2A Histone Family Member Z (H2AFZ) or Chromosome 1 Open Reading Frame 162 (C1 orf162).

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the non-human subject is a mammal.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the mammal is genetically-modified.

In some embodiments of the methods of the disclosure, the genetically-modified mammal is a model organism for the fibrotic lung disease.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the fibrotic lung disease is pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), an interstitial lung abnormality (ILA), or an asymptomatic ILA.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the fibrotic lung disease is pulmonary fibrosis or IPF.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the fibrotic lung disease is IPF.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the non-human subject carries a mutation in a sequence encoding MUC5B.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the mutation comprises a polymorphism in a sequence encoding a MUC5B promoter.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the polymorphism is rs35705950.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the non-human subject carries a mutation in a sequence encoding TERC, FAM13A, TERT, DSP, ZKSCAN1, AZGP1, OBFC1, MUC5B, AK025511, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the composition prevents the onset or development of a sign or symptom of the fibrotic lung disease.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the composition delays the onset or development of a sign or symptom of the fibrotic lung disease when compared to the expected onset of the a sign or symptom in the absence of treatment with the composition.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the composition delays the onset or development of a sign or symptom of the fibrotic lung disease when compared to the expected onset of the sign or symptom when treated using a standard therapeutic intervention.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the composition reduces the severity of a sign or symptom of the fibrotic lung disease when compared to the expected severity of the sign or symptom in the absence of treatment with the composition.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the composition reduces the severity of a sign or symptom of the fibrotic lung disease when compared to the expected severity of the sign or symptom when treated using a standard therapeutic intervention.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the standard therapeutic intervention comprises a N-acetylcysteine, pirfenidone, and nintedanib.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the standard therapeutic intervention comprises pirfenidone.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, an effective dosage of pirfenidone is about 2400 mg/day.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the effective dosage is administered orally as a capsule or a tablet.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the effective dosage is administered three times per day.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the effective dosage is administered according to an escalating dosage regimen.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the escalating dosage regimen comprises, administering to the non-human subject about 800 mg of pirfenidone per day for a first week; administering to the non-human subject about 1600 mg of pirfenidone per day for a second week; and administering to the non-human subject about 2400 mg of pirfenidone per day for the remainder of the treatment.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the escalating dosage regimen comprises, administering to the non-human subject a capsule or tablet comprising about 250 mg of pirfenidone three times a day for a first week; administering to the non-human subject two capsules or tablets comprising about 250 mg of pirfenidone three times a day for a second week; and administering to the non-human subject three capsules or tablets comprising about 250 mg of pirfenidone three times a day for the remainder of the treatment.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the capsule or tablet comprises 267 mg of pirfenidone.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the standard therapeutic intervention comprises nintedanib.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, an effective dosage of nintedanib is administered orally as a capsule or a tablet.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the effective dosage is about 300 mg/day.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the effective dosage is about 150 mg administered twice per day, wherein the daily doses are administered about 12 hours apart from one another.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the effective dosage is about 200 mg/day.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the effective dosage is about 100 mg administered twice per day, wherein the daily doses are administered about 12 hours apart from one another.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the non-human subject presents at least one sign of the fibrotic lung disease.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the at least one sign comprises gradual or unintended weight loss, clubbing of the fingers or toes, rapid and shallow breathing, fibrotic lesions in one or both lungs detectable by radiography, or a cough.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the compound prevents the onset of a secondary condition associated with a severe form of the fibrotic lung disease.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, the compound prevents the onset for at 1 year, 2 years, 3 years, 4 years, 5 years or any whole or fractional number of years in between.

In some embodiments of the methods of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure, secondary condition comprises a collapsed lung, an infected lung, a blood clot in a lung, lung cancer, respiratory failure, pulmonary hypertension, heart failure or death.

The disclosure provides a composition for the treatment of a fibrotic lung disease identified by a method of the disclosure, including, a method of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease of the disclosure.

The disclosure provides a method of treating fibrotic lung disease in a human subject of the disclosure comprising administering a therapeutically effective amount of a composition identified by a method of the disclosure, wherein the subject is asymptomatic and wherein the subject is at risk of developing the fibrotic lung disease. In some embodiments, the subject is wild type (e.g. does not comprises a mutation or a sequence variation) with respect to a nucleic acid or amino acid sequence encoding one or more of TERC, FAM13A, TERT, DSP, ZKSCAN1, AZGP1, OBFC1, MUC5B, AK025511, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7.

In some embodiments of the methods of treating fibrotic lung disease in a human subject of the disclosure by administering a composition identified by a method of the disclosure, the human subject presents radiographic Usual Interstitial Pneumonia (UIP).

In some embodiments of the methods of treating fibrotic lung disease in a human subject of the disclosure by administering a composition identified by a method of the disclosure, wherein the human subject has fibrotic interstitial lung disease (FILD).

In some embodiments of the methods of treating fibrotic lung disease in a human subject of the disclosure by administering a composition identified by a method of the disclosure, wherein the human subject has a blood relative with familial interstitial pneumonia (FIP).

In some embodiments of the methods of treating fibrotic lung disease in a human subject of the disclosure by administering a composition identified by a method of the disclosure, wherein the blood relative is a sibling.

In some embodiments of the methods of treating fibrotic lung disease in a human subject of the disclosure by administering a composition identified by a method of the disclosure, wherein the human subject has a mutation or a sequence variation in a nucleic acid or an amino acid sequence encoding TERC, FAM13A, TERT, DSP, ZKSCAN1, AZGP1, OBFC1, MUC5B, AK025511, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7.

In some embodiments of the methods of treating fibrotic lung disease in a human subject of the disclosure by administering a composition identified by a method of the disclosure, the mutation comprises a polymorphism in a sequence encoding a MUC5B promoter.

In some embodiments of the methods of treating fibrotic lung disease in a human subject of the disclosure by administering a composition identified by a method of the disclosure, the polymorphism is rs35705950.

In some embodiments of the methods of treating fibrotic lung disease in a human subject of the disclosure by administering a composition identified by a method of the disclosure, the fibrotic lung disease is pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), an interstitial lung abnormality (ILA), or an asymptomatic ILA.

In some embodiments of the methods of treating fibrotic lung disease in a human subject of the disclosure by administering a composition identified by a method of the disclosure, the fibrotic lung disease is pulmonary fibrosis or IPF.

In some embodiments of the methods of treating fibrotic lung disease in a human subject of the disclosure by administering a composition identified by a method of the disclosure, the fibrotic lung disease is IPF.

In some embodiments of the methods of treating fibrotic lung disease in a human subject of the disclosure by administering a composition identified by a method of the disclosure, the method prevents the onset of a secondary condition associated with a severe form of the fibrotic lung disease.

In some embodiments of the methods of treating fibrotic lung disease in a human subject of the disclosure by administering a composition identified by a method of the disclosure, a secondary condition comprises a collapsed lung, an infected lung, a blood clot in a lung, lung cancer, respiratory failure, pulmonary hypertension, heart failure or death.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

FIG. 1 is a map depicting an exemplary hierarchical clustering of differentially expressed genes for pre-pulmonary fibrosis subjects and normal subjects.

FIG. 2A-B is a pair of volcano plots showing serum sample quality control using Principal component analysis (PCA). FIG. 2A shows before outlier exclusion and FIG. 2B shows after outlier exclusion.

FIG. 3 is a volcano plot of 3315 plasma proteins, comparing results from 70 patients with established IPF and 70 controls. Solid red symbols represent 57 proteins that were significantly up-regulated and solid blue symbols 12 proteins that were significantly down-regulated in patients with IPF after controlling for multiple comparisons and age/gender/smoking.

FIG. 4 is a survival plot showing receiver operator curves of predictive model for PrePF in asymptomatic relatives from FIP families. Area Under Curve (AUC) values for each model are as follows: Gene Expression alone (red)=0.83, Clinical Predictors (blue)=0.87, Clinical Predictors+MUC5B genotype (green)=0.87, Clinical Predictors+Gene Expression Score (yellow)=0.95, Clinical Predictors+MUC5B genotype+Gene Expression Score (black)=0.95, indicating that a peripheral blood biomarker panel may improve the diagnostic power of a predictive model for PrePF in an at-risk population.

FIG. 5 is a graph showing MUC5B expression in IPF (N=203) and unaffected subjects (N=139) stratified by MUC5B promoter variant (rs35705950) genotype.

FIG. 6A is a microscopic image demonstrating that MUC5B is produced in bronchoalveolar epithelia of patients with IPF (brown staining in photomicrographs). Staining is increased in the airways of patients positive for rs35705950 (TT) compared to WT (GG).

FIG. 6B is a graph showing the percentage of MUC5B positive area of bronchiolar epithelium. Unbiased stereological assessment of staining demonstrates that the volume fraction of stained airways (% positive area) is significantly greater in both the GT heterozygotes and the TT homozygotes.

FIG. 7A-B is series of bar graphs showing that Scgb1a1- and SFPTC promoter show significant worsening of fibrosis (hydroxyproline) after bleomycin while Muc5b−/− mice are protected. FIG. 7A is a series of graphs and FIG. 7B is a series of confocal images showing that the concentration of Muc5b is directly related to the fibroproliferative response to bleomycin. Representative images from second harmonic generation (SHG) demonstrate increased lung collagen (red) in transgenic mice following bleomycin injury.

FIG. 8 is bar graph showing that the baseline expression of ER stress genes in lung tissue from WT and Scgb1a1 Muc5bTg mice. Muc5bTg mice have greater ER stress gene expression than their WT littermates (all genes in the ER stress pathway, with p<0.05). Bleomycin also induces ER stress (data not shown).

FIG. 9 is a pair of microscopic images showing enhanced CHOP (Ddit3) protein in wild type (WT, top photograph) and Scgb1a1-Muc5bTg mice (bottom photograph) after repeat bleomycin.

FIG. 10 is a pair of microscopic images and corresponding graphs showing the expanded mucus layer and decreased mucociliary transport in SFTPC-Muc5bTg mice compared to littermate wild-type mice. Statistical differences were assessed by Mann-Whitney U Test.

FIG. 11 is a series of schematic diagram showing that the MUC5B variant and other biomarkers can identify an at-risk population or those with PrePF, establishing the opportunity for primary and secondary prevention of IPF. The ‘at-risk’ population and the population with PrePF is large (19% with the MUC5B promoter variant and 1.8% of individuals ≥50 years of age respectively), IPF is diagnosed in a small population with established, end-stage disease and PrePF can be identified using the MUC5B variant rs35705950. Results indicate that PrePF (detected via chest CT scan) is associated with a poor prognosis suggesting that PrePF may be a harbinger of IPF.

FIG. 12 is a schematic diagram showing a method of screening at-risk populations (family members of patients with IPF) to identify individuals with PrePF. Focus is placed on identifying the genetic variants and biomarkers that increase the yield of PrePF on HRCT scan, in addition to gender, age, and physiology scores.

FIG. 13 is a table describing the baseline characteristics of patients with rheumatoid arthritis.

FIG. 14 is a table describing the genotypic association of MUC5B rs35705950 single nucleotide polymorphism in patients with RA, with and without interstitial lung disease

FIG. 15 is a table describing the dominant genotypic association of MUC5B rs35705950 single nucleotide polymorphism in patients with RA-ILD and a usual interstitial pneumonia or possible usual interstitial pneumonia pattern (RA-UIP) and in patients with RA-ILD and a pattern inconsistent with usual interstitial pneumonia (RA non-UIP).

FIG. 16A is a forest plot of odds ratios {OR) and 95% confidence intervals {C1) depicting the lack of association of the MUC5B rs35705950 promoter variant with RA without 1LD {RA-nolLD). The boxes indicate OR, and the horizontal lines indicate 95% C1 for the best-fitting genetic model for each association test. The black dotted line represents a mean OR value of 1. The red boxes and red lines indicate the overall OR and 95% C1, respectively. For comparisons between RA cases and controls, the associations were adjusted for the country of origin and sex. For intra-RA cases comparisons, the associations were adjusted for the country of origin, sex, age at inclusion and smoking.

FIG. 16B is a forest plot of odds ratios (OR) and 95% confidence intervals {C1) depicting the additive genotypic association of the MUC5B rs 35705950 promoter variant with RA-ILD. The red dotted line represent the mean value of overall OR value. The boxes indicate OR, and the horizontal lines indicate 95% C1 for the best-fitting genetic model for each association test. The black dotted line represents a mean OR value of 1. The red boxes and red lines indicate the overall OR and 95% C1, respectively. For comparisons between RA cases and controls, the associations were adjusted for the country of origin and sex. For intra-RA cases comparisons, the associations were adjusted for the country of origin, sex, age at inclusion and smoking.

FIG. 16C is a forest plot of odds ratios {OR) and 95% confidence intervals {C1) depicting dominant genotypic association of the MUC5B re35705950 promoter variant with ILD among patients with RA and those with the usual interstitial pneumonia or possible usual interstitial pneumonia (UIP) pattern. The boxes indicate OR, and the horizontal lines indicate 95% C1 for the best-fitting genetic model for each association test. The red dotted line represent the mean value of overall OR value. The black dotted line represents a mean OR value of 1. The red boxes and red lines indicate the overall OR and 95% C1, respectively. For comparisons between RA cases and controls, the associations were adjusted for the country of origin and sex. For intra-RA cases comparisons, the associations were adjusted for the country of origin, sex, age at inclusion and smoking.

FIG. 17 is a series of photographs depicting MUC5B expression in explanted lung issue from rheumatoid arthritis associates interstitial lung disease. Representative lung tissue images from unaffected control (GG genotype, Panel A), RA-ILD case #1 (GG genotype, Panel B), and RA-ILD case #2 (GT genotype, Panel C). Low power views with high power view insets identified. Panel A—low power view of normal lung; top and middle insets with high power view of bronchiole with MUC5B staining; bottom inset with high power view of alveolar epithelia. Panel B and C—low power view of the usual interstitial pneumonia pattern in explanted lung tissue of RA-ILD; top inset with high power view of bronchiole with MUC5B staining; middle and bottom insets with high power view of MUC5B staining in metaplastic epithelia lining honeycomb cysts and MUC5B staining of mucous in honeycomb cysts.

FIG. 18 is a flow chart depicting the screening and enrollment process for study subjects.

FIG. 19A-D is a series of photographs depicting High-resolution CT (HRCT) images of: 19A) chest from a study subject whose scan was read as normal, without signs of interstitial lung disease or fibrosis. 19B) HRCT image from subject who was categorized as having “Probable Fibrotic ILD.” 19C) Representative HRCT image from subject who was characterized as having “Definite Fibrotic ILD.” 19D) HRCT image from a case of previously diagnosed, established Idiopathic Pulmonary Fibrosis (IPF) in one of the study families.

FIG. 20 is a table depicting a summary of characteristics of study subjects used in quantitative CT Analyses.

FIG. 21A-F is a series of photographs depicting representative axial HRCT images visually assessed as “No Fibrosis” (21A), “Probable Fibrotic ILD” (21C) and “Definite Fibrotic ILD” (E). Below each is the corresponding quantitative HRCT results for the above scan: (21B) “No Fibrosis” fibrosis extent 1.7% (fibrosis score=0.55), (21D) “Probable Fibrotic ILD” fibrosis extent 18.5% (fibrosis score 2.92), (F) “Definite Fibrotic ILD” fibrosis extent 35.5% (fibrosis score 3.60), Classification results color coded as follows: green=normal lung, blue=airway, yellow=reticular abnormality, magenta=ground glass opacity, red=honeycombing.

FIG. 22 is a table depicting Screening Cohort Subject Characteristics. * DNA available on a total of 489 subjects (404 No Fibrosis and 75 PrePF subjects). ** Odds ratios reported in this table were calculated from a mixed effects logistic regression model including age (as a continuous variable), male sex, ever smoker (yes/no), and MUC5B promoter variant (rs35705950) genotype. ***In the reported model, rs35705950 coded as a dominant allele; in log-additive genetic model, p=0.05, as well.

FIG. 23 is a table depicting patterns of CT abnormalities in scans with probable or definite fibrotic ILD. * Because a confident single diagnosis was relatively uncommon, most cases included consideration of several patterns. For this reason, the percentages add up to more than 100%.

FIG. 24 is a box plot depicting fibrosis score by visual diagnosis. Boxplots of fibrosis scores based on quantitative HRCT assessment for each visual diagnosis category. Fibrosis score means were significantly different (ANOVA, p<0.0001) across groups defined by visual diagnosis. Comparison of fibrosis score between groups showed significant differences for all comparisons (p<0.01 for all).

FIG. 25A-C is a series of graphs depicting Receiver Operating Characteristic (ROC) curves for quantitative imaging measures of Fibrosis and PrePF. FIG. 5A depicts ROC curves for visual diagnosis compared to log HAA scores. FIG. 5B depicts ROC Curves for visual diagnosis compared to fibrosis scores. ROC analysis showed that fibrosis score discriminates subjects with visual diagnosis of PrePF. Average area under the curve (AUC) in fivefold cross validation was 0.85 (range 0.83-0.87) and average accuracy, sensitivity, and specificity in the test partitions were 0.83 (range 0.74-0.86), 0.74 (range 0.56-0.92) and 0.84 (range 0.76-0.89) respectively. Optimal threshold for fibrosis score ranged from 1.40-1.42.

FIG. 5C depicts Density plots of fibrosis scores for visually diagnosed PrePF (pink) and No Fibrosis (blue) scans—the fibrosis score optimal threshold is indicated with the red line (1.40).

FIG. 26 is a series of tables depicting Dyspnea questionnaire data. FIG. 26A depicts breathlessness responses for the cohort. FIG. 26B depicts breathlessness responses by Visual CT diagnosis.

FIG. 27 is a graph that depicts the prevalence of PrePF in FIP Siblings Cohort by Age and MUC5B Genotype. PrePF prevalence in this FIP siblings cohort increases by age, as shown in this graph. By age >60 years, the prevalence of PrePF differed significantly based on MUC5B genotype (*p=0.02). Subjects with the variant are depicted by the red line, while those without it are depicted with the blue line.

FIG. 28 is a table depicting subject characteristics based on Quantitative Fibrosis Score. Clinical characteristics and genotype breakdown of subjects with quantitative HRCT analyses. The cutoff of 1.4 for the logarithm of fibrosis score is based on analyses presented in the text. * p-value compares characteristic between groups. Linear regression values regress fibrosis score on age, male sex, smoking history, and MUC5B promoter variant. **In the reported model, rs35705950 coded as a dominant allele given small number of TT subjects.

FIG. 29 is a table depicting an exploratory genetic association study of 13 pulmonary fibrosis susceptibility variants in RA-ILD.

DETAILED DESCRIPTION OF THE DISCLOSURE

The present disclosure provides a method of treating a fibrotic lung disease in a subject comprising administering to the subject an effective amount of a therapeutic agent, wherein the subject is asymptomatic and wherein the subject is at risk of developing the fibrotic lung disease.

Methods of Identifying a Therapeutic Agent of the Disclosure or Target Thereof

The disclosure provides a method of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease, comprising administering to a non-human subject a dose of a composition that modifies transcription or translation of a sequence encoding Mucin 5B (MUC5B), Telomerase RNA Component (TERC), Family with sequence similarity 13 member A (FAM13A), Telomerase Reverse Transcriptase (TERT), Desmoplakin (DSP), Zinc-alpha 2-Glycoprotein 1 (AZGP1), Oligonucleotide/oligosaccharide-binding Fold Containing 1 (OBFC1), ATPase Phospholipid Transporting 11A (ATP11A), Isovaleryl-CoA dehydrogenase (IVD)/Dispatched RND Transporter Family Member 2 (DISP2), Dipeptidyl Peptidase 9 (DPP9), Sialic Acid Binding Ig-Like Lectin 14 (SIGLEC14), Adrenomedullin 2 (ADM2), Tetraspanin 5 (TSPAN5), Calcium/Calmodulin-Dependent Protein Kinase Kinase 1 (CAMKK1) or Matrix Metalloprotease-7 (MMP-7), wherein the dose of the composition is tolerable to the non-human subject and wherein the dose of the composition is therapeutically effective.

The disclosure provides method of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease, comprising administering to a non-human subject a composition that modifies an activity of a product of a sequence encoding MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7, wherein the dose of the composition is tolerable to the non-human subject and wherein the dose of the composition is therapeutically effective.

In some embodiments of the methods of the disclosure, the composition that modifies transcription or translation decreases or inhibits transcription or translation. In some embodiments, the composition decreases or inhibits transcription or translation of a sequence encoding a gene selected from the group consisting of Leukotriene A4 Hydrolase (LTA4H), Surfactant Protein B (SFTPB), Breast Cancer Anti-Estrogen Resistance 3 (BCAR3), C—X—C motif Chemokine Ligand 13 (CXCL13), EPH Receptor A2 (EPHA2), Serum Amyloid A1 (SAA1), Phospholipase A2 Group IIA (PLA2G2A), Insulin-Like Growth Factor Binding Protein 3 (IGFBP3), C-C Motif Chemokine Ligand 28 (CCL28), S100 Calcium Binding Protein A12 (S100A12), Thromboxane A Synthase 1 (TBXAS1), Leukocyte Cell Derived Chemotaxin 1 (LECT1), Complement C3 (C3), Gastrin Releasing Peptide (GRP), C-Reactive Protein (CRP), Vitrin (VIT), Insulin-Like Growth Factor Binding Protein 1 (IGFBP1), Family with Sequence Similarity 173 Member A (FAM173A), Natriuretic Peptide A (NPPA), Secreted Frizzled Related Protein 1 (SFRP1), Ezrin (EZR), Inter-Alpha-Trypsin Inhibitor Heavy Chain Family Member 5 (ITIH5), Pleckstrin and Sec7 Domain Containing 2 (PSD2), Galectin 3 Binding Protein (LGALS3BP), Catenin Beta 1 (CTNNB1), Chromodomain Y Like 2 (CDYL2), Matrix Metallopeptidase 7 (MMPI), Apolipoprotein B (APOB), Proline and Arginine Rich End Leucine Rich Repeat Protein (PRELP), Eukaryotic Translation Initiation Factor 1A, X-linked (EIF1AX), Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF), TNF Receptor Superfamily Member 13C (TNFRSF13C), Deformed Epidermal Autoregulatory Factor 1 transcription factor (DEAF1), Tumor Protein Translationally-Controlled 1 (TPT1), Unc-5 Netrin Receptor B (UNC5B), Phosphatidylethanolamine Binding Protein 1 (PEBP1), Syntaxin 8 (STX8), Polymeric Immunoglobulin Receptor (PIGR), Adenine Phosphoribosyltransferase (APRT), Matrix Metallopeptidase 3 (MMP3), Galectin 7 (LGALS7), Bruton Tyrosine Kinase (BTK), NSFL1 Cofactor (NSFL1C), FER Tyrosine Kinase (FER), Regenerating Family Member 1 Beta (REG1B), SMAD Family Member 2 (SMAD2), Interleukin 1 Receptor Like 1 (IL1RL1), C-C Motif Chemokine Ligand 18 (CCL18), Acid Phosphatase 2 Lysosomal (ACP2), Eukaryotic Translation Initiation Factor 4E Family Member 2 (EIF4E2), Neurexin 3 (NRXN3), IGF Like Family Member 1 (IGFL1), NME/NM23 Nucleoside Diphosphate Kinase 1 (NME1), Potassium Voltage-Gated Channel Isk-Related Family Member 1-Like (KCNE1L) or Neurexophilin 2 (NXPH2).

In some embodiments of the methods of the disclosure, the composition that modifies transcription or translation increases or activates transcription or translation. In some embodiments, the composition increases or activates transcription or translation of a sequence encoding a gene selected from the group consisting of Surfactant Protein D (SFTPD), Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), Histone Cluster 1 H1 Family Member C (HIST1H1C), YTH Domain Containing 1 (YTHDC1), Plexin A1 (PLXNA1), Serine Peptidase Inhibitor Kazal Type 6 (SPINK6), LDL Receptor Related Protein Associated Protein 1 (LRPAP1), Secretoglobin Family 3A Member 1 (SCGB3A1), H2A Histone Family Member Z (H2AFZ) or Chromosome 1 Open Reading Frame 162 (C1 orf162).

In some embodiments of the methods of the disclosure, the composition that modifies an activity decreases or inhibits the activity. In some embodiments, the composition decreases or inhibits the activity of a sequence encoding a gene selected from Leukotriene A4 Hydrolase (LTA4H), Surfactant Protein B (SFTPB), Breast Cancer Anti-Estrogen Resistance 3 (BCAR3), C-X-C motif Chemokine Ligand 13 (CXCL13), EPH Receptor A2 (EPHA2), Serum Amyloid A1 (SAA1), Phospholipase A2 Group IIA (PLA2G2A), Insulin-Like Growth Factor Binding Protein 3 (IGFBP3), C-C Motif Chemokine Ligand 28 (CCL28), 5100 Calcium Binding Protein A12 (S100A12), Thromboxane A Synthase 1 (TBXAS1), Leukocyte Cell Derived Chemotaxin 1 (LECT1), Complement C3 (C3), Gastrin Releasing Peptide (GRP), C-Reactive Protein (CRP), Vitrin (VIT), Insulin-Like Growth Factor Binding Protein 1 (IGFBP1), Family with Sequence Similarity 173 Member A (FAM173A), Natriuretic Peptide A (NPPA), Secreted Frizzled Related Protein 1 (SFRP1), Ezrin (EZR), Inter-Alpha-Trypsin Inhibitor Heavy Chain Family Member 5 (ITIH5), Pleckstrin and Sec7 Domain Containing 2 (PSD2), Galectin 3 Binding Protein (LGALS3BP), Catenin Beta 1 (CTNNB1), Chromodomain Y Like 2 (CDYL2), Matrix Metallopeptidase 7 (MMPI), Apolipoprotein B (APOB), Proline and Arginine Rich End Leucine Rich Repeat Protein (PRELP), Eukaryotic Translation Initiation Factor 1A, X-linked (EIF1AX), Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF), TNF Receptor Superfamily Member 13C (TNFRSF13C), Deformed Epidermal Autoregulatory Factor 1 transcription factor (DEAF1), Tumor Protein Translationally-Controlled 1 (TPT1), Unc-5 Netrin Receptor B (UNC5B), Phosphatidylethanolamine Binding Protein 1 (PEBP1), Syntaxin 8 (STX8), Polymeric Immunoglobulin Receptor (PIGR), Adenine Phosphoribosyltransferase (APRT), Matrix Metallopeptidase 3 (MMP3), Galectin 7 (LGALS7), Bruton Tyrosine Kinase (BTK), NSFL1 Cofactor (NSFL1C), FER Tyrosine Kinase (FER), Regenerating Family Member 1 Beta (REG1B), SMAD Family Member 2 (SMAD2), Interleukin 1 Receptor Like 1 (IL1RL1), C-C Motif Chemokine Ligand 18 (CCL18), Acid Phosphatase 2 Lysosomal (ACP2), Eukaryotic Translation Initiation Factor 4E Family Member 2 (EIF4E2), Neurexin 3 (NRXN3), IGF Like Family Member 1 (IGFL1), NME/NM23 Nucleoside Diphosphate Kinase 1 (NME1), Potassium Voltage-Gated Channel Isk-Related Family Member 1-Like (KCNE1L) or Neurexophilin 2 (NXPH2).

In some embodiments of the methods of the disclosure, the composition that modifies an activity increases or activates the activity. In some embodiments, the composition increases or activates the activity of a sequence encoding Surfactant Protein D (SFTPD), Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), Histone Cluster 1 H1 Family Member C (HIST1H1C), YTH Domain Containing 1 (YTHDC1), Plexin A1 (PLXNA1), Serine Peptidase Inhibitor Kazal Type 6 (SPINK6), LDL Receptor Related Protein Associated Protein 1 (LRPAP1), Secretoglobin Family 3A Member 1 (SCGB3A1), H2A Histone Family Member Z (H2AFZ) or Chromosome 1 Open Reading Frame 162 (C1 orf162).

In some embodiments of the methods of the disclosure, the non-human subject is a mammal. In some embodiments, mammal is genetically-modified. In some embodiments, the genetically-modified mammal is a model organism for the fibrotic lung disease.

In some embodiments of the methods of the disclosure, the fibrotic lung disease is pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), an interstitial lung abnormality (ILA), or an asymptomatic ILA. In some embodiments, the fibrotic lung disease is pulmonary fibrosis or IPF. In some embodiments, the fibrotic lung disease is IPF.

In some embodiments of the methods of the disclosure, the non-human subject carries a mutation in a sequence encoding MUC5B. In some embodiments, the mutation comprises a polymorphism in a sequence encoding a MUC5B promoter. In some embodiments, the polymorphism is rs35705950. Alternatively, or in addition, in some embodiments, the non-human subject carries a mutation in a sequence encoding TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7.

In some embodiments of the methods of the disclosure, the composition prevents the onset or development of a sign or symptom of the fibrotic lung disease.

In some embodiments of the methods of the disclosure, the composition delays the onset or development of a sign or symptom of the fibrotic lung disease when compared to the expected onset of the sign or symptom in the absence of treatment with the composition. In some embodiments, the composition delays the onset or development of a sign or symptom of the fibrotic lung disease when compared to the expected onset of the sign or symptom when treated using a standard therapeutic intervention.

In some embodiments of the methods of the disclosure, the composition reduces the severity of a sign or symptom of the fibrotic lung disease when compared to the expected severity of the sign or symptom in the absence of treatment with the composition. In some embodiments, the composition reduces the severity of a sign or symptom of the fibrotic lung disease when compared to the expected severity of the sign or symptom when treated using a standard therapeutic intervention.

In some embodiments of the methods of the disclosure, the standard therapeutic intervention comprises a N-acetylcysteine, pirfenidone, and nintedanib.

In some embodiments of the methods of the disclosure, the standard therapeutic intervention comprises pirfenidone. In some embodiments, an effective dosage of pirfenidone is about 2400 mg/day. In some embodiments, the effective dosage is administered orally as a capsule or a tablet. In some embodiments, the effective dosage is administered three times per day. In some embodiments, the effective dosage is administered according to an escalating dosage regimen. In some embodiments, the escalating dosage regimen comprises (a) administering to the non-human subject about 800 mg of pirfenidone per day for a first week; (b) administering to the non-human subject about 1600 mg of pirfenidone per day for a second week; and (c) administering to the non-human subject about 2400 mg of pirfenidone per day for the remainder of the treatment. In some embodiments, the escalating dosage regimen comprises (a) administering to the non-human subject a capsule or tablet comprising about 250 mg of pirfenidone three times a day for a first week; (b) administering to the non-human subject two capsules or tablets comprising about 250 mg of pirfenidone three times a day for a second week; and (c) administering to the non-human subject three capsules or tablets comprising about 250 mg of pirfenidone three times a day for the remainder of the treatment. In some embodiments, the capsule or tablet comprises 267 mg of pirfenidone.

In some embodiments of the methods of the disclosure, the standard therapeutic intervention comprises nintedanib. In some embodiments, an effective dosage of nintedanib is administered orally as a capsule or a tablet. In some embodiments, the effective dosage is about 300 mg/day. In some embodiments, the effective dosage is about 150 mg administered twice per day, wherein the daily doses are administered about 12 hours apart from one another. In some embodiments, the effective dosage is about 200 mg/day. In some embodiments, the effective dosage is about 100 mg administered twice per day, wherein the daily doses are administered about 12 hours apart from one another.

In some embodiments of the methods of the disclosure, the non-human subject presents at least one sign of the fibrotic lung disease. In some embodiments, the at least one sign comprises gradual or unintended weight loss, clubbing of the fingers or toes, rapid and shallow breathing, fibrotic lesions in one or both lungs detectable by radiography, or a cough.

In some embodiments of the methods of the disclosure, the compound prevents the onset of a secondary condition associated with a severe form of the fibrotic lung disease. In some embodiments, the compound prevents the onset for at 1 year, 2 years, 3 years, 4 years, 5 years or any whole or fractional number of years in between. In some embodiments, the secondary condition comprises a collapsed lung, an infected lung, a blood clot in a lung, lung cancer, respiratory failure, pulmonary hypertension, heart failure or death.

The disclosure provides a composition for the treatment of a fibrotic lung disease identified by a method of the disclosure for identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease.

Subjects of the Disclosure

The disclosure provides a method of treating a fibrotic lung disease in a human subject comprising administering to the subject the composition for the treatment of a fibrotic lung disease identified by a method of the disclosure for identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease, wherein the subject is asymptomatic and wherein the subject is at risk of developing the fibrotic lung disease.

In some embodiments of the methods of treating a fibrotic lung disease in a human subject of the disclosure, the human subject presents radiographic Usual Interstitial Pneumonia (UIP). In some embodiments, the human subject has fibrotic interstitial lung disease (FILD). In some embodiments, the human subject has a blood relative with familial interstitial pneumonia (FIP). In some embodiments, the blood relative is a sibling. Alternatively, or in addition, in some embodiments, the human subject has a mutation in a sequence encoding MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7. In some embodiments, the mutation comprises a polymorphism in a sequence encoding a MUC5B promoter. In some embodiments, the polymorphism is rs35705950.

In some embodiments of the methods of treating a fibrotic lung disease in a human subject of the disclosure, the fibrotic lung disease is pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), an interstitial lung abnormality (ILA), or an asymptomatic ILA. In some embodiments, the fibrotic lung disease is pulmonary fibrosis or IPF. In some embodiments, the fibrotic lung disease is IPF.

In some embodiments of the methods of treating a fibrotic lung disease in a human subject of the disclosure, the method prevents the onset of a secondary condition associated with a severe form of the fibrotic lung disease. In some embodiments, the secondary condition comprises a collapsed lung, an infected lung, a blood clot in a lung, lung cancer, respiratory failure, pulmonary hypertension, heart failure or death.

Idiopathic Pulmonary Fibrosis (IPF)

IPF is localized to the lung and is characterized by a pattern of heterogeneous, subpleural patches of fibrotic, remodeled lung, and often results in death within 3-5 years of diagnosis. IPF affects 5 million people worldwide, disproportionately affects men, is associated with cigarette smoking, increases with age, is inexplicably increasing in prevalence, and is likely underdiagnosed. Most patients with IPF are discovered in the advanced stage when little can be done to influence survival. There is a critical unmet need in idiopathic pulmonary fibrosis (IPF) for an early detection and prevention of IPF. Earlier diagnosis of IPF detects subjects with a lower burden of fibrotic lung disease providing an opportunity for secondary prevention of this progressive disease and changes the clinical approach to patients with IPF from palliative to preventive.

Early detection and prevention of idiopathic pulmonary fibrosis (IPF) is critical. As demonstrated herein, treatment of subjects at risk for developing PrePF is based on two central concepts of first, understanding that PrePF is essential for primary and secondary prevention of IPF and second, that similar to asymptomatic family members of familial IPF (FIP; ≥2 family members with IPF), asymptomatic family members of sporadic IPF represent an at-risk population for PrePF. These central concepts are supported by the observation that 1) IPF has a pre-symptomatic phase and PrePF appears to be a harbinger of IPF, 2) familial and sporadic IPF are similar etiologically, 3) MUC5B promoter variant is critical to early disease recognition and 4) identification of PrePF represents an opportunity to prevent extensive lung fibrosis. As shown herein, a common gain-of-function MUC5B promoter variant rs35705950 is a strong risk factor (genetic and otherwise), accounting for at least 30% of the total risk of developing IPF. The MUC5B promoter variant rs35705950 may be used to identify individuals with PrePF. MUC5B promoter variant rs35705950 is also predictive of radiographic progression of PrePF and is present in over 50% of non-Hispanic white patients with IPF and is also associated with unique clinical and biological IPF phenotypes. PrePF can be predicted using a combination of clinical risk factors, the MUC5B promoter variant rs35705950, and a panel of biomarkers. This disclosure provides methods of treating subjects with Preclinical Pulmonary Fibrosis (PrePF) and who may also be at risk for developing IPF. The methods of the disclosure fundmentally change the clinical approach to treating subjects with IPF, shifting the focus from a merely palliative to a proactive and preventive therapy.

Rheumatoid Arthritis-Associated Interstitial Lung Disease (RA-ILD)

Rheumatoid arthritis (RA) is a common inflammatory and autoimmune disease that is associated with progressive impairment, systemic complications and increased mortality. Interstitial lung disease (RA-ILD) is detected in up to 60% of patients with RA on high-resolution computed-tomography (HRCT), is clinically significant in 10%, and is a leading cause of morbidity and mortality in patients with RA.

RA-ILD shares several characteristics with idiopathic pulmonary fibrosis (IPF), including common environmental risk factors, the high prevalence of the usual interstitial pneumonia (UIP) pattern, the progressive nature of the disease, and poor survival. The hypothesis of a shared genetic background between IPF and RA-ILD was recently suggested by a whole-exome sequencing (WES) genetic association study in patients with RA-ILD, revealing an excess of mutations in genes in RA-ILD previously associated with familial interstitial pneumonia (FIP) including TERT, RTEL1, PARN and SFTPC.

The common gain-of-function promoter variant rs3570595013 of the gene encoding mucin5B (MUC5B) is the strongest genetic risk factor for IPF, observed in at least 50% of the cases of IPF and accounting for 30% of the risk of developing this disease. The MUC5B promoter variant is associated with increased expression of MUC5B in lung parenchyma of unaffected controls and cases of IPF. Consequently, it is hypothesized that the MUC5B promoter variant rs35705950 would also contribute to the occurrence of RA-ILD. To test this hypothesis, a multi-ethnic association study of the MUC5B promoter variantand RA-ILD in seven distinct case series was performed.

The MUC5B promoter variant rs35705950, the strongest genetic risk factor for IPF, is also a strong risk factor for RA-ILD, especially among those with radiographic evidence of UIP. Of note, the effect of the MUC5B promoter variant on the development of ILD associated with RA was similar in magnitude and direction to that observed in IPF.

The relationship between the MUC5B promoter variant and RA-ILD may be specific to UIP and may not generalizable to other autoimmune conditions of the lung. The MUC5B promoter variant has not been found to be associated with risk of ILDs linked to systemic sclerosis or autoimmune myositis. Unlike these other types of ILD, RA-ILD shares more characteristics with IPF, notably the increased frequency of the UIP pattern (both radiologic and histologic), an increased prevalence of male sex and older age, and genetic susceptibility as assessed by an excess of mutations in genes linked to FIP in a cohort of RA-ILD, and now the MUC5B promoter variant rs35705950.

The disclosure demonstrates that the MUC5B promoter variant is a risk factor for UIP, and not simply limited to IPF and RA-ILD. In fact, emerging studies have identified the MUC5B promoter variant as a risk factor for chronic hypersensitivity pneumonitis, another condition known to have a sub-phenotype of UIP. Further, since HRCT underestimates the presence of ILD and the UIP pattern of fibrosis, our point estimates for association with the MUC5B variant are likely conservative. Similar to IPF, early forms of RA-ILD can be identified using the MUC5B promoter variant as biomarker.

The disclosure demonstrates that Muc5b is overexpressed by the bronchoalveolar epithelia and MUC5B mRNA is co-expressed by cells expressing surfactant protein C, as has been shown in IPF. These findings suggest either type 2 alveolar epithelial cells can express MUC5B or that in patients with RA-ILD, the cells in the distal airspace de-differentiate. Importantly, the disclosure demonstrates for the first time that cells that overexpress MUC5B are undergoing ER stress, a recognized mechanism of cell injury and repair. In aggregate, these findings indicate that the gain-of-function MUC5B promoter variant rs35705950 injures alveolar epithelia by inducing ER stress.

RA-ILD is a complex genetic phenotype with the minor allele of the MUC5B promoter variant rs35705950 identified as a risk factor for the disease. The odds ratios for the association of MUC5B promoter variant with RA-ILD is equivalent to that observed with IPF and substantively higher than those for the most other common risk variants for RA-ILD, including cigarette smoking and the human leukocyte antigen locus for RA.

The MUC5B promoter variant is a risk factor for UIP in general and may prove relevant beyond RA-ILD and IPF.

Expression of MUC5B in the bronchoalveolar epithelia co-incident with markers of ER stress suggest that the MUC5B promoter variant may be causing pulmonary fibrosis by initiating microscopic foci of injury and repair.

The MUC5B promoter variant appears to predict ILD in the RA population, identifying potential opportunities for early ILD detection in patients with RA.

Preclinical Idiopathic Pulmonary Fibrosis

Better understanding and recognition of early pulmonary fibrosis is critical because medical therapies have been shown to slow progression, not to reverse or even stabilize established fibrosis—therefore, intervention before irreversible fibrosis has become extensive has the potential to improve quality of life and decrease morbidity. While IPF affects approximately 5 million people worldwide, between 1.8 and 14% of the general population ≥50 years of age have radiologic findings of undiagnosed pulmonary fibrosis. Large cohort studies indicate that interstitial lung abnormalities, postulated to represent early pulmonary fibrosis, are associated with increased mortality, and that most of these abnormalities progress over time. Members of families with 2 or more cases of pulmonary fibrosis (FIP, Familial Interstitial Pneumonia) have been identified as an “at-risk” population. In a previous study of FIP relatives, 14% had interstitial lung abnormalities on high resolution computed tomography (HRCT), and 35% had an abnormal transbronchial biopsy indicating interstitial lung disease.

HRCT provides visualization of the lung parenchyma and plays a key role in the diagnosis of the Idiopathic Interstitial Pneumonias (IIPs), including IPF. Currently, visual diagnosis by thoracic radiologists, in conjunction with multidisciplinary clinical conference, is the gold standard for diagnosing IIPs. However, visual assessment is imprecise and hampered by inter-observer variation. Quantitative HRCT (qHRCT) evaluation provides measures of fibrosis extent that, in subjects diagnosed with IPF, correlate with degree of physiologic impairment at baseline, and may be more sensitive to subtle changes in disease status than routinely used physiological metrics. The design and utility of quantitative methods in the context of early forms of fibrotic ILD requires further study. Deep learning methods have been increasingly used in imaging to identify and classify CT patterns, and may be particularly valuable in detection of early lung fibrosis.

PrePF is prevalent among FIP relatives, and a texture-based quantitative method of HRCT analyses is useful in identifying these abnormalities in this population, and key risk factors, including the MUC5B promoter variant, predict those at risk of this disease. PrePF subjects are older, more likely to be male, and more likely to have smoked than the unaffected subjects; additionally, the gain-of-function MUC5B promoter variant rs35705950, which has been shown in prior studies to be associated with pulmonary fibrosis, is more common in PrePF subjects when compared to their unaffected family members. Given the subtlety of the fibrotic change in many of these cases of PrePF, the high prevalence of potential UIP pattern on HRCT scan suggests that PrePF subjects may progress to IPF over time.

Methods for Detecting a Genetic Variant

The present disclosure also provides methods of detecting the biomarkers of the present disclosure. Methods of detecting a genetic variant are further described in US Application US 2016-0060701A1(the contents of which are incorporated herein by reference in their entirety). The practice of the present disclosure employs, unless otherwise indicated, conventional methods of analytical biochemistry, microbiology, molecular biology and recombinant DNA techniques within the skill of the art. Such techniques are explained fully in the literature. (See, e.g., Sambrook, J. et al. Molecular Cloning: A Laboratory Manual. 3rd, ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N. Y., 2000; DNA Cloning: A Practical Approach, Vol. I & II (D. Glover, ed.); Oligonucleotide Synthesis (N. Gait, ed., Current Edition); Nucleic Acid Hybridization (B. Hames & S. Higgins, eds., Current Edition); Transcription and Translation (B. Hames & S. Higgins, eds., Current Edition); CRC Handbook of Parvoviruses, Vol. I & II (P. Tijessen, ed.); Fundamental Virology, 2nd Edition, Vol. I & II (B. N. Fields and D. M. Knipe, eds.)).

The methods of the invention are not limited to any particular way of detecting the presence or absence of a genetic variant (e.g. SNP) and can employ any suitable method to detect the presence or absence of a variant(s), of which numerous detection methods are known in the art. Dynamic allele-specific hybridization (DASH) can be used to detect a genetic variant. DASH genotyping takes advantage of the differences in the melting temperature in DNA that results from the instability of mismatched base pairs. The process can be vastly automated and encompasses a few simple principles. Thus, the aspects and embodiments described herein provide methods for assessing the presence or absence of SNPs in a sample (e.g. biological sample) from a subject suspected of having or developing an interstitial lung disease (e.g., because of family history). In certain embodiments, one or more SNPs are screened in one or more samples from a subject. The SNPs can be associated with one or more genes, e.g., one or more genes or other genes associated with mucous secretions as disclosed herein.

Typically, the target genomic segment is amplified and separated from non-target sequence, e.g., through use of a biotinylated primer and chromatography. A probe that is specific for the particular allele is added to the amplification product. The probe can be designed to hybridize specifically to a variant sequence or to the dominant allelic sequence. The probe can be either labeled with or added in the presence of a molecule that fluoresces when bound to double-stranded DNA. The signal intensity is then measured as temperature is increased until the Tm can be determined. A non-matching sequence (either genetic variant or dominant allelic sequence, depending on probe design), will result in a lower than expected Tm.

DASH genotyping relies on a quantifiable change in Tm, and is thus capable of measuring many types of mutations, not just SNPs. Other benefits of DASH include its ability to work with label free probes and its simple design and performance conditions.

Molecular beacons can also be used to detect a genetic variant. This method makes use of a specifically engineered single-stranded oligonucleotide probe. The oligonucleotide is designed such that there are complementary regions at each end and a probe sequence located in between. This design allows the probe to take on a hairpin, or stem-loop, structure in its natural, isolated state. Attached to one end of the probe is a fluorophore and to the other end a fluorescence quencher. Because of the stem-loop structure of the probe, the fluorophore is in close proximity to the quencher, thus preventing the molecule from emitting any fluorescence. The molecule is also engineered such that only the probe sequence is complementary to the targeted genomic DNA sequence.

If the probe sequence of the molecular beacon encounters its target genomic DNA sequence during the assay, it will anneal and hybridize. Because of the length of the probe sequence, the hairpin segment of the probe will be denatured in favor of forming a longer, more stable probe-target hybrid. This conformational change permits the fluorophore and quencher to be free of their tight proximity due to the hairpin association, allowing the molecule to fluoresce.

If on the other hand, the probe sequence encounters a target sequence with as little as one non-complementary nucleotide, the molecular beacon will preferentially stay in its natural hairpin state and no fluorescence will be observed, as the fluorophore remains quenched. The unique design of these molecular beacons allows for a simple diagnostic assay to identify SNPs at a given location. If a molecular beacon is designed to match a wild-type allele and another to match a mutant of the allele, the two can be used to identify the genotype of an individual. If only the first probe's fluorophore wavelength is detected during the assay then the individual is homozygous to the wild type. If only the second probe's wavelength is detected then the individual is homozygous to the mutant allele. Finally, if both wavelengths are detected, then both molecular beacons must be hybridizing to their complements and thus the individual must contain both alleles and be heterozygous.

A microarray can also be used to detect genetic variants. Hundreds of thousands of probes can be arrayed on a small chip, allowing for many genetic variants or SNPs to be interrogated simultaneously. Because SNP alleles only differ in one nucleotide and because it is difficult to achieve optimal hybridization conditions for all probes on the array, the target DNA has the potential to hybridize to mismatched probes. This can be addressed by using several redundant probes to interrogate each SNP. Probes can be designed to have the SNP site in several different locations as well as containing mismatches to the SNP allele. By comparing the differential amount of hybridization of the target DNA to each of these redundant probes, it is possible to determine specific homozygous and heterozygous alleles.

Restriction fragment length polymorphism (RFLP) can be used to detect genetic variants and SNPs. RFLP makes use of the many different restriction endonucleases and their high affinity to unique and specific restriction sites. By performing a digestion on a genomic sample and determining fragment lengths through a gel assay it is possible to ascertain whether or not the enzymes cut the expected restriction sites. A failure to cut the genomic sample results in an identifiably larger than expected fragment implying that there is a mutation at the point of the restriction site which is rendering it protected from nuclease activity.

PCR- and amplification-based methods can be used to detect genetic variants. For example, tetra-primer PCR employs two pairs of primers to amplify two alleles in one PCR reaction. The primers are designed such that the two primer pairs overlap at a SNP location but each matches perfectly to only one of the possible alleles. As a result, if a given allele is present in the PCR reaction, the primer pair specific to that allele will produce product but not the alternative allele with a different allelic sequence. The two primer pairs can be designed such that their PCR products are of a significantly different length allowing for easily distinguishable bands by gel electrophoresis, or such that they are differently labeled.

Primer extension can also be used to detect genetic variants. Primer extension first involves the hybridization of a probe to the bases immediately upstream of the SNP nucleotide followed by a ‘mini-sequencing’ reaction, in which DNA polymerase extends the hybridized primer by adding a base that is complementary to the SNP nucleotide. The incorporated base that is detected determines the presence or absence of the SNP allele. Because primer extension is based on the highly accurate DNA polymerase enzyme, the method is generally very reliable. Primer extension is able to genotype most SNPs under very similar reaction conditions making it also highly flexible. The primer extension method is used in a number of assay formats, and can be detected using e.g., fluorescent labels or mass spectrometry.

Primer extension can involve incorporation of either fluorescently labeled ddNTP or fluorescently labeled deoxynucleotides (dNTP). With ddNTPs, probes hybridize to the target DNA immediately upstream of SNP nucleotide, and a single, ddNTP complementary to the SNP allele is added to the 3′ end of the probe (the missing 3′-hydroxyl in didioxynucleotide prevents further nucleotides from being added). Each ddNTP is labeled with a different fluorescent signal allowing for the detection of all four alleles in the same reaction. With dNTPs, allele-specific probes have 3′ bases which are complementary to each of the SNP alleles being interrogated. If the target DNA contains an allele complementary to the 3′ base of the probe, the target DNA will completely hybridize to the probe, allowing DNA polymerase to extend from the 3′ end of the probe. This is detected by the incorporation of the fluorescently labeled dNTPs onto the end of the probe. If the target DNA does not contain an allele complementary to the probe's 3′ base, the target DNA will produce a mismatch at the 3′ end of the probe and DNA polymerase will not be able to extend from the 3′ end of the probe.

The iPLEX® SNP genotyping method takes a slightly different approach, and relies on detection by mass spectrometer. Extension probes are designed in such a way that many different SNP assays can be amplified and analyzed in a PCR cocktail. The extension reaction uses ddNTPs as above, but the detection of the SNP allele is dependent on the actual mass of the extension product and not on a fluorescent molecule. This method is for low to medium high throughput, and is not intended for whole genome scanning.

Primer extension methods are, however, amenable to high throughput analysis. Primer extension probes can be arrayed on slides allowing for many SNPs to be genotyped at once. Broadly referred to as arrayed primer extension (APEX), this technology has several benefits over methods based on differential hybridization of probes. Comparatively, APEX methods have greater discriminating power than methods using differential hybridization, as it is often impossible to obtain the optimal hybridization conditions for the thousands of probes on DNA microarrays (usually this is addressed by having highly redundant probes).

Oligonucleotide ligation assays can also be used to detect genetic variants. DNA ligase catalyzes the ligation of the 3′ end of a DNA fragment to the 5′ end of a directly adjacent DNA fragment. This mechanism can be used to interrogate a SNP by hybridizing two probes directly over the SNP polymorphic site, whereby ligation can occur if the probes are identical to the target DNA. For example, two probes can be designed; an allele-specific probe which hybridizes to the target DNA so that its 3′ base is situated directly over the SNP nucleotide and a second probe that hybridizes the template upstream (downstream in the complementary strand) of the SNP polymorphic site providing a 5′ end for the ligation reaction. If the allele-specific probe matches the target DNA, it will fully hybridize to the target DNA and ligation can occur. Ligation does not generally occur in the presence of a mismatched 3′ base. Ligated or unligated products can be detected by gel electrophoresis, MALDI-TOF mass spectrometry or by capillary electrophoresis.

The 5′-nuclease activity of Taq DNA polymerase can be used for detecting genetic variants. The assay is performed concurrently with a PCR reaction and the results can be read in real-time. The assay requires forward and reverse PCR primers that will amplify a region that includes the SNP polymorphic site. Allele discrimination is achieved using FRET, and one or two allele-specific probes that hybridize to the SNP polymorphic site. The probes have a fluorophore linked to their 5′ end and a quencher molecule linked to their 3′ end. While the probe is intact, the quencher will remain in close proximity to the fluorophore, eliminating the fluorophore's signal. During the PCR amplification step, if the allele-specific probe is perfectly complementary to the SNP allele, it will bind to the target DNA strand and then get degraded by 5′-nuclease activity of the Taq polymerase as it extends the DNA from the PCR primers. The degradation of the probe results in the separation of the fluorophore from the quencher molecule, generating a detectable signal. If the allele-specific probe is not perfectly complementary, it will have lower melting temperature and not bind as efficiently. This prevents the nuclease from acting on the probe.

Førster resonance energy transfer (FRET) detection can be used for detection in primer extension and ligation reactions where the two labels are brought into close proximity to each other. It can also be used in the 5′-nuclease reaction, the molecular beacon reaction, and the invasive cleavage reactions where the neighboring donor/acceptor pair is separated by cleavage or disruption of the stem-loop structure that holds them together. FRET occurs when two conditions are met. First, the emission spectrum of the fluorescent donor dye must overlap with the excitation wavelength of the acceptor dye. Second, the two dyes must be in close proximity to each other because energy transfer drops off quickly with distance. The proximity requirement is what makes FRET a good detection method for a number of allelic discrimination mechanisms.

A variety of dyes can be used for FRET, and are known in the art. The most common ones are fluorescein, cyanine dyes (Cy3 to Cy7), rhodamine dyes (e.g. rhodamine 6G), the Alexa series of dyes (Alexa 405 to Alexa 730). Some of these dyes have been used in FRET networks (with multiple donors and acceptors). Optics for imaging all of these require detection from UV to near IR (e.g. Alex 405 to Cy7), and the Atto series of dyes (Atto-Tec GmbH). The Alexa series of dyes from Invitrogen cover the whole spectral range. They are very bright and photostable.

Example dye pairs for FRET labeling include Alexa-405/Alex-488, Alexa-488/Alexa-546, Alexa-532/Alexa-594, Alexa-594/Alexa-680, Alexa-594/Alexa-700, Alexa-700/Alexa-790, Cy3/Cy5, Cy3.5/Cy5.5, and Rhodamine-Green/Rhodamine-Red, etc. Fluorescent metal nanoparticles such as silver and gold nanoclusters can also be used (Richards et al. (2008) J Am Chem Soc 130:5038-39; Vosch et al. (2007) Proc Natl Acad Sci USA 104:12616-21; Petty and Dickson (2003) J Am Chem Soc 125:7780-81 Available filters, dichroics, multichroic mirrors and lasers can affect the choice of dye.

In Vitro Complexes

Provided herein are nucleic acid complexes, e.g., formed in in vitro assays to indicate the presence of a genetic variant sequence. One of skill will understand that a nucleic acid complex can also be formed to detect the presence of a dominant allelic sequence, depending on the design of the probe or primer, e.g., in assays to distinguish homozygous and heterozygous subjects.

In some embodiments, the complex comprises a first nucleic acid hybridized to a genetic variant nucleic acid, wherein the genetic variant nucleic acid is a genetic variant in a gene selected from MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7. In some embodiments, the genetic variant nucleic acid is an amplification product. In some embodiments, the genetic variant nucleic acid is on genomic DNA, e.g., from a subject that has or is suspected of having an interstitial lung disease. In some embodiments, the first nucleic acid is an amplification product or a primer extension product. In some embodiments, the first nucleic acid is labeled. In some embodiments, the nucleic acid complex further comprises a second nucleic acid hybridized to the genetic variant nucleic acid. In some embodiments, the second nucleic acid is labeled e.g., with a FRET or other fluorescent label. In some embodiments, the first and second nucleic acids form a FRET pair when hybridized to a genetic variant sequence.

In some embodiments, the nucleic acid complex further comprises an enzyme, such as a DNA polymerase (e.g., standard DNA polymerase or thermostable polymerase such as Taq) or ligase.

The present disclosure includes but is not limited to the following embodiments:

A method for determining if an individual is predicted to develop and/or progress rapidly with an interstitial pneumonia comprising: detecting in a biological sample from the individual, at least one of: a) the presence of a marker polymorphism selected from the group consisting of: rs35705950; and/or, b) a level of gene expression of a marker gene or plurality of marker genes selected from the group consisting of: a marker gene having at least 95% sequence identity with at least one sequence selected from the group consisting of MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7, or homologs or variants thereof c) polypeptides encoded by the marker genes of b) d) fragments of polypeptides of c); and e) a polynucleotide which is fully complementary to at least a portion of a marker gene of b); wherein the presence of the plurality of markers is indicative of whether an individual will develop a disease. In some embodiments, the genes detected share 100% sequence identity with the corresponding marker gene in b). In some embodiments, the presence or level of at least one of the plurality of markers is determined and compared to a standard level or reference set. In some embodiments, the standard level or reference set is determined according to a statistical procedure for risk prediction. In some embodiments, the statistical procedure for risk prediction comprises using the sum of the gene expression of the marker or markers or the presence or absence of a set of markers, weighted by a Proportional Hazards coefficient. In some embodiments, the presence of the at least one marker is determined by detecting the presence or absence or expression level of a polypeptide. In some embodiments, the method further comprises detecting the presence of the polypeptide using a reagent that specifically binds to the polypeptide or a fragment thereof. In some embodiments, the reagent is selected from the group consisting of an antibody, an antibody derivative, and an antibody fragment. In some embodiments, the presence of the marker is determined by obtaining the sequence of genomic DNA at the locus of the polymorphism. In some embodiments, the presence of the marker is determined by obtaining RNA from the biological sample; generating cDNA from the RNA; amplifying the cDNA with probes or primers for marker genes; obtaining from the amplified cDNA the expression levels of the genes or gene expression products in the sample. In some embodiments, the individual is a human.

In some embodiments, the method further comprises: a) comparing the expression level of the marker gene or plurality of marker genes in the biological sample to a control level of the marker gene(s) selected from the group consisting of: a control level of the marker gene that has been correlated with interstitial lung disease, the risk of developing interstitial lung disease, or having a interstitial lung disease; and a control level of the marker that has been correlated with slow or no progression of interstitial lung disease, or low risk of developing an interstitial lung disease; and b) selecting the individual as being predicted to progress rapidly in the development of interstitial pneumonia, if the expression level of the marker gene in the individual's biological sample is statistically similar to, or greater than, the control level of expression of the marker gene that has been correlated with interstitial lung disease, or c) selecting the individual as being predicted to not develop interstitial lung disease, or to progress slowly, if the level of the marker gene in the individual's biological sample is statistically less than the control level of the marker gene that has been correlated with interstitial lung disease.

In some embodiments, the method further comparing the presence of a polymorphism, in the biological sample to a set of genetic variants or polymorphic markers from an individual or control group having developed interstitial lung disease, and, selecting the individual as being predicted to develop or to progress with interstitial pneumonia if the polymorphic markers present in the biological sample are identical to or statistically similar to a set of polymorphic markers from the individual or control group or, selecting the individual as being predicted to develop or rapidly progress with interstitial pneumonia, if the polymorphic markers present in the biological sample are not identical to or statistically similar to the set of genetic variants or polymorphic markers from the individual or control group.

A method for monitoring the progression of interstitial lung disease in a subject, comprising: i) measuring expression levels of a plurality of gene markers in a first biological sample obtained from the subject, wherein the plurality of markers comprise a plurality of markers selected from the group consisting of: a marker gene having at least 95% sequence identity with a sequence selected from the group consisting of a) MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7, or homologs or variants thereof; b) polypeptides encoded by the marker genes of a), c) fragments of polypeptides of d); and e) a polynucleotide which is fully complementary to at least a portion of a marker gene of b); ii) measuring expression levels of the plurality of markers in a second biological sample obtained from the subject; and iii) comparing the expression level of the marker measured in the first sample with the level of the marker measured in the second sample. In some embodiments, the marker genes detected share 100% sequence identity with the corresponding marker gene in a). In some embodiments, the method further comprises performing a follow-up step selected from the group consisting of CT scan of the chest and pathological examination of lung tissues from the subject. In some embodiments, the first biological sample from the subject is obtained at a time to, and the second biological sample from the subject is obtained at a later time t₁. In some embodiments, the first biological sample and the second biological sample are obtained from the subject are obtained more than once over a range of times.

A method of assessing the efficacy of a treatment for interstitial lung disease or interstitial pneumonia in a subject, the method comprising comparing: i) the expression level of a marker measured in a first sample obtained from the subject at a time to, wherein the marker is selected from the group consisting of a) a marker gene having at least 95% sequence identity with a sequence selected from the group consisting of MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7, or homologs or variants thereof; b) polypeptides encoded by the marker genes of a)

c) fragments of polypeptides of b); and d) a polynucleotide which is fully complementary to at least a portion of a marker gene of a); ii) the level of the marker in a second sample obtained from the subject at time t₁; and, iii) performing a follow-up step selected from CT scan of the chest and pathological examination of lung tissues from the subject; wherein a decrease in the level of the marker in the second sample relative to the first sample is an indication that the treatment is efficacious for treating interstitial pneumonia in the subject. In some embodiments, the genes detected share 100% sequence identity with the corresponding marker gene in a). In some embodiments, the time t0 is before the treatment has been administered to the subject, and the time t1 is after the treatment has been administered to the subject. In some embodiments, the comparing is repeated over a range of times.

An assay system for predicting individual prognosis therapy for interstitial pneumonia comprising a means to detect at least one of: a) the presence of a marker polymorphism selected from the group consisting of: rs35705950; and/or, b) a level of gene expression of a marker gene or plurality of marker genes selected from the group consisting of: a marker gene having at least 95% sequence identity with a sequence selected from the group consisting of MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7, or homologs or variants thereof c) polypeptides encoded by the marker genes of b) d) fragments of polypeptides of c); and e) a polynucleotide which is fully complementary to at least a portion of a marker gene of b). In some embodiments, the means to detect comprises nucleic acid probes comprising at least 10 to 50 contiguous nucleic acids of the marker polymorphisms or gene(s), or complementary nucleic acid sequences thereof. In some embodiments, the means to detect comprises binding ligands that specifically detect polypeptides encoded by the marker genes. In some embodiments, the genes detected share 100% sequence identity with the corresponding marker gene in b). In some embodiments, the means to detect comprises at least one of nucleic acid probe and binding ligands disposed on an assay surface. In some embodiments, the assay surface comprises a chip, array, or fluidity card. In some embodiments, the probes comprise complementary nucleic acid sequences to at least 10 to 50 nucleic acid sequences of the marker genes. In some embodiments, the binding ligands comprise antibodies or binding fragments thereof. In some embodiments, the assay system further comprises: a control selected from information containing a predetermined control level or set of genetic variants or polymorphic markers that has been correlated with diagnosis, development, progression, or life expectancy in interstitial lung disease patients.

A method of detecting a level of gene expression of one or more marker genes in a human subject with interstitial pneumonia, comprising, optionally, obtaining a biological sample from a human individual with interstitial pneumonia; detecting the level of expression of a gene selected from MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7, or homologs or variants thereof, in one or more cells from the biological sample from the individual. In some embodiments, the method further comprises detecting the level of expression of a gene selected from MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7, or homologs or variants thereof, in one or more cells from the biological sample from the individual. In some embodiments, the method further comprises detecting the level of expression of a gene selected from MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7, or homologs or variants thereof in one or more cells from the biological sample from the individual.

A method of treating an interstitial lung disease in a subject in need of such treatment, comprising: detecting a level of one or more marker genes selected from MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7, or homologs or variants thereof in a biological sample obtained from the human subject; and, administering an effective amount of an effective treatment. In some embodiments, the method further comprises detecting the level of expression of a gene selected from MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7, or homologs or variants thereof, in one or more cells from the biological sample from the individual. In some embodiments, the method further comprises detecting the level of expression of a gene selected from MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7, or homologs or variants thereof, in one or more cells from the biological sample from the individual.

Detection of Genetic Variants

Methods of detecting a genetic variant are further described, for example, in U.S. Pat. No. 8,673,565 (the contents of which are herein incorporated by reference in their entirety). Genetic variations in the mucin genes are associated with pulmonary diseases. These genetic variations can be found in any part of the gene, e.g., in the regulatory regions, introns, or exons. Relevant genetic variations may also be found the intergene regions, e.g., in sequences between mucin genes. Insertions, substitutions, and deletions are included in genetic variants. Single nucleotide polymorphisms (SNPs) are exemplary genetic variants.

In particular, 14 independent SNPs are associated with pulmonary disorders (e.g. FIP or IPF). The studies disclosed herein demonstrate that presence of one or more of these SNPs associated with MUC5B can lead to predisposition to a pulmonary disorder. In addition, in some embodiments, if present, some of these SNPs are related to a transcription factor binding site. The transcription factor binding site can effect modulation of MUC5B expression, for example E2F3 loss, and HOXA9 and PAX-2 generation.

The disclosure thus provides methods for assessing the presence or absence of SNPs in a sample from a subject suspected of having or developing a pulmonary disorder (e.g., because of family history). In certain embodiments, one or more SNPs are screened in one or more samples from a subject. The SNPs can be associated with one or more genes, e.g., one or more MUC genes or other genes associated with mucous secretion. In some embodiments, a MUC gene associated SNP is associated with MUC5B and/or another MUC gene, such as MUC5AC or MUC1. SNPs contemplated for diagnostic, treatment, or prognosis can include SNPs found within a MUC gene and/or within a regulatory or promoter region associated with a MUC gene. For example, one or more SNPs can include, but are not limited to, detection of the SNPs of MUC5B alone or in combination with other genetic variations or SNPs and/or other diagnostic or prognostic methods.

Methods for detecting genetic variants such as a SNP are known in the art, e.g., Southern or Northern blot, nucleotide array, amplification methods, etc. Primers or probes are designed to hybridize to a target sequence. For example, genomic DNA can be screened for the presence of an identified genetic element of using a probe based upon one or more sequences, e.g., using a probe with substantial identity to a subsequence of the MUC5B gene. Expressed RNA can also be screened, but may not include all relevant genetic variations. Various degrees of stringency of hybridization may be employed in the assay. As the conditions for hybridization become more stringent, there must be a greater degree of complementarity between the probe and the target for duplex formation to occur. Thus, high stringency conditions are typically used for detecting a SNP.

Thus, in some embodiments, a genetic variant MUC5B gene in a subject is detected by contacting a nucleic acid in a sample from the subject with a probe having substantial identity to a subsequence of the MUC5B gene, and determining whether the nucleic acid indicates that the subject has a genetic variant MUC5B gene. In some cases, the sample can be processed prior to amplification, e.g., to separate genomic DNA from other sample components. In some cases, the probe has at least 90, 92, 94, 95, 96, 98, 99, or 100% identity to the MUC5B gene subsequence. Typically, the probe is between 10-500 nucleotides in length, e.g., 10-100, 10-40, 10-20, 20-100, 100-400, etc. In the case of detecting a SNP, the probe can be even shorter, e.g., 8-20 nucleotides in length. In some cases, the MUC5B gene sequence to be detected includes at least 8 contiguous nucleotides, e.g., at least 10, 15, 20, 25, 30, 35 or more contiguous nucleotides. In some embodiments, the sequence to be detected includes 8 contiguous nucleotides, e.g., at least 10, 15, 20, 25, 30, 35 or more contiguous nucleotides.

The degree of stringency can be controlled by temperature, ionic strength, pH and/or the presence of a partially denaturing solvent such as formamide. For example, the stringency of hybridization is conveniently varied by changing the concentration of formamide within the range up to and about 50%. The degree of complementarity (sequence identity) required for detectable binding will vary in accordance with the stringency of the hybridization medium and/or wash medium. In certain embodiments, in particular for detection of a particular SNP, the degree of complementarity is about 100 percent. In other embodiments, sequence variations can result in <100% complementarity, <90% complimentarity probes, <80% complimentarity probes, etc., in particular, in a sequence that does not involve a SNP. In some examples, e.g., detection of species homologs, primers may be compensated for by reducing the stringency of the hybridization and/or wash medium.

High stringency conditions for nucleic acid hybridization are well known in the art. For example, conditions may comprise low salt and/or high temperature conditions, such as provided by about 0.02 M to about 0.15 M NaCl at temperatures of about 50° C. to about 70° C. Other exemplary conditions are disclosed in the following Examples. It is understood that the temperature and ionic strength of a desired stringency are determined in part by the length of the particular nucleic acid(s), the length and nucleotide content of the target sequence(s), the charge composition of the nucleic acid(s), and by the presence or concentration of formamide, tetramethylammonium chloride or other solvent(s) in a hybridization mixture. Nucleic acids can be completely complementary to a target sequence or exhibit one or more mismatches.

Nucleic acids of interest can also be amplified using a variety of known amplification techniques. For instance, polymerase chain reaction (PCR) technology may be used to amplify target sequences (e.g., genetic variants) directly from DNA, RNA, or cDNA. In some embodiments, a stretch of nucleic acids is amplified using primers on either side of a targeted genetic variation, and the amplification product is then sequenced to detect the targeted genetic variation (using, e.g., Sanger sequencing, Pyrosequencing, Nextgen® sequencing technologies). For example, the primers can be designed to hybridize to either side of the upstream regulatory region of the MUC5B gene, and the intervening sequence determined to detect a SNP in the promoter region. In some embodiments, one of the primers can be designed to hybridize to the targeted genetic variant. In some cases, a genetic variant nucleotide can be identified using RT-PCR, e.g., using labeled nucleotide monomers. In this way, the identity of the nucleotide at a given position can be detected as it is added to the polymerizing nucleic acid. The Scorpion™ system is a commercially available example of this technology.

Thus, in some embodiments, a genetic variant MUC5B gene in a subject is detected by amplifying a nucleic acid in a sample from the subject to form an amplification product, and determining whether the amplification product indicates a genetic variant MUC5B gene. In some cases, the sample can be processed prior to amplification, e.g., to separate genomic DNA from other sample components. In some cases, amplifying comprises contacting the sample with amplification primers having substantial identity to MUC5B genomic subsequences, e.g., at least 90, 92, 94, 95, 96, 98, 99, or 100% identity. Typically, the sequence to be amplified is between 30-1000 nucleotides in length, e.g., 50-500, 50-400, 100-400, 50-200, 100-300, etc. In some cases, the sequence to be amplified or detected includes at least 8 contiguous nucleotides, e.g., at least 10, 15, 20, 25, 30, 35 or more contiguous nucleotides. In some embodiments, the sequence to be amplified or detected includes 8 contiguous nucleotides, e.g., at least 10, 15, 20, 25, 30, 35 or more contiguous nucleotides. In some aspects, the contiguous nucleotides include nucleotide 28.

Amplification techniques can also be useful for cloning nucleic acid sequences, to make nucleic acids to use as probes for detecting the presence of a target nucleic acid in samples, for nucleic acid sequencing, for control samples, or for other purposes. Probes and primers are also readily available from commercial sources, e.g., from Invitrogen, Clonetech, etc.

Detection of Expression Levels

Expression of a given gene, e.g., MUC5B or another mucin, pulmonary disease marker, or standard (control), is typically detected by detecting the amount of RNA (e.g., mRNA) or protein. Sample levels can be compared to a control level.

Methods for detecting RNA are largely cumulative with the nucleic acid detection assays described above. RNA to be detected can include mRNA. In some embodiments, a reverse transcriptase reaction is carried out and the targeted sequence is then amplified using standard PCR. Quantitative PCR (qPCR) or real time PCR (RT-PCR) is useful for determining relative expression levels, when compared to a control. Quantitative PCR techniques and platforms are known in the art, and commercially available (see, e.g., the qPCR Symposium website, available at qpersymposium.com). Nucleic acid arrays are also useful for detecting nucleic acid expression. Customizable arrays are available from, e.g., Affimatrix. An exemplary human MUC5B mRNA sequence, e.g., for probe and primer design, can be found at GenBank Accession No. AF086604.1.

Protein levels can be detected using antibodies or antibody fragments specific for that protein, natural ligands, small molecules, aptamers, etc. An exemplary human MUC5B sequence, e.g., for screening a targeting agent, can be found at UniProt Accession No. 000446.

Antibody based techniques are known in the art, and described, e.g., in Harlow & Lane (1988) Antibodies: A Laboratory Manual and Harlow (1998) Using Antibodies: A Laboratory Manual; Wild, The Immunoassay Handbook, 3d edition (2005) and Law, Immunoassay: A Practical Guide (1996). The assay can be directed to detection of a molecular target (e.g., protein or antigen), or a cell, tissue, biological sample, liquid sample or surface suspected of carrying an antibody or antibody target.

A non-exhaustive list of immunoassays includes: competitive and non-competitive formats, enzyme linked immunosorption assays (ELISA), microspot assays, Western blots, gel filtration and chromatography, immunochromatography, immunohistochemistry, flow cytometry or fluorescence activated cell sorting (FACS), microarrays, and more. Such techniques can also be used in situ, ex vivo, or in vivo, e.g., for diagnostic imaging.

Aptamers are nucleic acids that are designed to bind to a wide variety of targets in a non-Watson Crick manner. An aptamer can thus be used to detect or otherwise target nearly any molecule of interest, including a pulmonary disease associated protein. Methods of constructing and determining the binding characteristics of aptamers are well known in the art. For example, such techniques are described in U.S. Pat. Nos. 5,582,981, 5,595,877 and 5,637,459. Aptamers are typically at least 5 nucleotides, 10, 20, 30 or 40 nucleotides in length, and can be composed of modified nucleic acids to improve stability. Flanking sequences can be added for structural stability, e.g., to form 3-dimensional structures in the aptamer.

Protein detection agents described herein can also be used as a treatment and/or diagnosis of pulmonary disease or predictor of disease progression, e.g., propensity for survival, in a subject having or suspected of developing a pulmonary disorder. In certain embodiments, MUC5B antibodies can be used to assess MUC5B protein levels in a subject having or suspected of developing a pulmonary disorder. It is contemplated herein that antibodies or antibody fragments may be used to modulate MUC5B production in a subject having or suspected of developing a pulmonary disease. In certain embodiments, one or more agents capable of modulating MUC5B may be used to treat a subject having or suspected of developing a pulmonary disorder. One or more antibodies or antibody fragments may be generated to detect one or more of the SNPs disclosed herein by any method known in the art.

In certain embodiments, MUC5B diagnostic tests may include, but are not limited to, alone or in combination, analysis of rs35705950 SNP in MUC5B gene, MUC5B mRNA levels, and/or MUC5B protein levels.

Additional Pulmonary Disease Markers

The above methods of detection can be applied to additional pulmonary disease markers. That is, the expression level or presence of genetic variants of at least one additional pulmonary disease marker gene can be determined, or the activity of the marker protein can be determined, and compared to a standard control for the pulmonary disease marker. The examination of additional pulmonary disease markers can be used to confirm a diagnosis of pulmonary disease, monitor disease progression, or determine the efficacy of a course of treatment in a subject.

In some cases, pulmonary disease is indicated by an increased number of lymphocytes, e.g., CD4+CD28− cells.

Genetic variations in the following genes are associated with pulmonary disease: Surfactant Protein A2, Surfactant Protein B, Surfactant Protein C, TERC, TERT, IL-1RN, IL-1α, IL-1β, TNF, Lymphotoxin a, TNF-RII, IL-10, IL-6, IL-12, IFNγ, TGFβ, CR1, ACE, IL-8, CXCR1, CXCR2, MUC1 (KL6), or MUC5AC. Thus, the invention further includes methods of determining whether the genome of a subject comprises a genetic variant of at least one gene selected from these genes. The presence of a genetic variant indicates that the subject has or is at risk of developing pulmonary disease. Said determining can optionally be combined with determining whether the genome of the subject comprises a genetic variant MUC5B gene, or determining whether the subject has an elevated level of MUC5B RNA or protein to confirm or strengthen the diagnosis or prognosis.

Abnormal expression in the following genes can also be indicative of pulmonary disease: Surfactant Protein A, Surfactant Protein D, KL-6/MUC1, CC16, CK-19, Ca 19-9, SLX, MCP-1, MIP-1a, ITAC, glutathione, type III procollagen peptide, sIL-2R, ACE, neopterin, beta-glucuronidase, LDH, CCL-18, CCL-2, CXCL12, MMPI, and osteopontin. Thus, the expression of one of these genes can be detected and compared to a control, wherein an abnormal expression level indicates that the subject has or is at risk of developing pulmonary disease. Said determining can optionally be combined with determining whether the genome of the subject comprises a genetic variant MUC5B gene, or determining whether the subject has an elevated level of MUC5B RNA or protein to confirm or strengthen the diagnosis or prognosis.

Biomarkers

The present disclosure provides a peripheral blood biomarker profile for IPF to demonstrate the use of a predictive biomarker profile in cases of preclinical pulmonary fibrosis (PrePF) derived from families with familial IPF. The present disclosure also provides biomarker identification for association between each genetic, epigenetic or protein (gene product) biomarker with PrePF and the predictive value of the combination of biomarkers associated with PrePF.

A large cohort of families with familial IPF for genetic research was established, including 937 families with ≥2 cases of IPF, and 2375 family members that have been previously phenotyped as unaffected. This study focuses on subjects with PrePF to elucidate the processes active in early disease pathogenesis and to predict or prevent the irreversible fibroproliferative process. Genetic risk factors, especially the MUC5B promoter variant, identifies individuals with preclinical interstitial changes on chest CT scan that progress and are associated with reduced survival. Biomarkers may be used to identify those subjects with PrePF among those at-risk for IPF. Given the irreversible nature of IPF, even approved treatments (pirfenidone and nintedanib) only modestly slow progression and have not been shown to alter the 3-5 year survival. Pirfenidone and nintedanib are effective in patients with mild disease, suggesting that patients with PrePF may be targeted for early intervention, before most of the lung has been irreversibly remodeled.

Table 1 below shows additional gene expression changes present in subjects with IPF compared to controls. Specifically, the expression of the genes listed in Table 1 are upregulated in IPF compared to the expression of these same genes in control subjects. Accordingly, the discovery of elevated expression levels of one or more genes listed in Table 1 compared to a control in an asymptomatic subject may indicate that the subject has PrePF and/or that the subject is at risk for developing IPF.

In some embodiments of the methods of the disclosure, the subject has a mutation in a nucleic acid or amino acid sequence encoding a gene or gene product that is upregulated in a subject having a fibrotic pulmonary disease of the disclosure. In some embodiments of the methods of the disclosure, the subject has a mutation in a nucleic acid or amino acid sequence encoding Leukotriene A4 Hydrolase (LTA4H), Surfactant Protein B (SFTPB), Breast Cancer Anti-Estrogen Resistance 3 (BCAR3), C-X-C motif Chemokine Ligand 13 (CXCL13), EPH Receptor A2 (EPHA2), Serum Amyloid A1 (SAA1), Phospholipase A2 Group IIA (PLA2G2A), Insulin-Like Growth Factor Binding Protein 3 (IGFBP3), C-C Motif Chemokine Ligand 28 (CCL28), 5100 Calcium Binding Protein A12 (S100A12), Thromboxane A Synthase 1 (TBXAS1), Leukocyte Cell Derived Chemotaxin 1 (LECT1), Complement C3 (C3), Gastrin Releasing Peptide (GRP), C-Reactive Protein (CRP), Vitrin (VIT), Insulin-Like Growth Factor Binding Protein 1 (IGFBP1), Family with Sequence Similarity 173 Member A (FAM173A), Natriuretic Peptide A (NPPA), Secreted Frizzled Related Protein 1 (SFRP1), Ezrin (EZR), Inter-Alpha-Trypsin Inhibitor Heavy Chain Family Member 5 (ITIH5), Pleckstrin and Sec7 Domain Containing 2 (PSD2), Galectin 3 Binding Protein (LGALS3BP), Catenin Beta 1 (CTNNB1), Chromodomain Y Like 2 (CDYL2), Matrix Metallopeptidase 7 (MMPI), Apolipoprotein B (APOB), Proline and Arginine Rich End Leucine Rich Repeat Protein (PRELP), Eukaryotic Translation Initiation Factor 1A, X-linked (EIF1AX), Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF), TNF Receptor Superfamily Member 13C (TNFRSF13C), Deformed Epidermal Autoregulatory Factor 1 transcription factor (DEAF1), Tumor Protein Translationally-Controlled 1 (TPT1), Unc-5 Netrin Receptor B (UNCSB), Phosphatidylethanolamine Binding Protein 1 (PEBP1), Syntaxin 8 (STX8), Polymeric Immunoglobulin Receptor (PIGR), Adenine Phosphoribosyltransferase (APRT), Matrix Metallopeptidase 3 (MMP3), Galectin 7 (LGALS7), Bruton Tyrosine Kinase (BTK), NSFL1 Cofactor (NSFL1C), FER Tyrosine Kinase (FER), Regenerating Family Member 1 Beta (REG1B), SMAD Family Member 2 (SMAD2), Interleukin 1 Receptor Like 1 (IL1RL1), C-C Motif Chemokine Ligand 18 (CCL18), Acid Phosphatase 2 Lysosomal (ACP2), Eukaryotic Translation Initiation Factor 4E Family Member 2 (EIF4E2), Neurexin 3 (NRXN3), IGF Like Family Member 1 (IGFL1), NME/NM23 Nucleoside Diphosphate Kinase 1 (NME1), Potassium Voltage-Gated Channel Isk-Related Family Member 1-Like (KCNE1L) or Neurexophilin 2 (NXPH2).

TABLE 1 TARGET_GENE_SYM- ORGAN- B-H Fold BOL ISM p-value q-value Change LTA4H Human 8.70E−43 3.13E−39 3.912 SFTP8 Human 1.17E−37 2.10E−34 3.399 BCAR3 Human 4.28E−25 3.85E−22 2.906 CXCL13 Human 1.30E−29 1.56E−26 2.904 EPHA2 Human 9.62E−23 6.93E−20 2.651 SAA1 Human 6.01E−07 7.84E−06 2.631 PLA2GZA Human 8.19E−21 2.95E−18 2.171 Igfbp3 Mouse 1.18E−18 2.66E−16 2.149 CCL28 Human 1.22E−22 7.30E−20 2.135 S100A12 Human 1.06E−20 3.45E−18 2.125 TBXAS1 Human 1.60E−21 7.20E−19 2.11 LECT1 Human 4.17E−19 1.00E−16 2.082 C3 Human 7.08E−07 8.95E−06 2.062 GRP Human 8.35E−09 1.66E−07 1.988 CSP Human 1.36E−08 2.61E−07 1.957 VIT Human 2.47E−17 4.45E−15 1.929 IGFBP1 Human 4.32E−11 1.56E−09 1.914 FAM173A Human 2.19E−13 1.84E−11 1.904 NPPA Human 5.02E−12 2.58E−10 1.877 SFRP1 Human 1.74E−20 5.23E−18 1.866 EZR Human 6.41E−10 1.72E−08 1.809 ITIH5 Human 5.11E−21 2.04E−18 1.705 PSD2 Human 5.38E−18 1.08E−15 1.689 LGAL538P Human 8.06E−22 4.15E−19 1.678 1.18E−05 0.000102 1.668 CTNNB1 Human 5.66E−12 2.87E−10 1.625 CDVL2 Human 4.11E−07 5.59E−06 1.622 MMP7 Human 1.56E−19 4.02E−17 1.621 APOB Human 8.73E−13 6.42E−11 1.597 PRELP Human 1.13E−10 3.53E−09 1.595 EIF1AX Human 2.13E−06 2.31E−05 1.59 MANF Human 0.00458  0.015006 1.585 TNFRSF13C Human 1.77E−11 7.31E−10 1.573 C3 Human 2.40E−16 3.93E−14 1.566 DEAF1 Human 0.000221 0.001192 1.565 TPT1 Human 1.22E−12 7.82E−11 1.548 UNC5B Human 2.06E−34 2.18E−12 1.547 PEBP1 Human 4.92E−11 1.72E−09 1.544 STX8 Human 8.82E−12 4.13E−10 1.537 PIGR Human 1.29E−09 3.19E−08 1.532 APRT Human 1.51E−07 2.26E−06 1.525 MMP3 Human 9.50E−07 1.15E−05 1.524 LGAL57 Human 7.51E−05 0.000474 1.514 BTK Human 1.47E−09 3.52E−08 1.511 NSFL1C Human 7.33E−11 2.40E−09 1.506 FER Human 2.24E−07 3.24E−06 1.503 REG1B Human 6.68E−11 2.25E−09 1.502 SMAD2 Human 4.39E−10 1.25E−08 1.493 IL1RL1 Human 9.55E−07 1.15E−05 1.492 CCL18 Human 1.25E−13 1.07E−11 1.491 ACP2 Human 3.73E−08 6.33E−07 1.488 EIF4E2 Human 1.67E−12 1.02E−10 1.483 NRXN3 Human 2.33E−17 4.42E−15 1.48 IGFL1 Human 5.07E−10 1.40E−08 1.474 NME1 Human 1.43E−10 4.39E−09 1.463 KCNE1L Human 3.93E−20 1.09E−17 1.462 NXPH2 Human 9.66E−30 2.47E−08 1.451

Table 2 below shows additional gene expression changes present in subjects with IPF compared to controls. Specifically, the expression of the genes listed in Table 2 are downregulated in IPF compared to the expression of these same genes in control subjects. Accordingly, the discovery of decreased expression levels of one or more genes listed in Table 2 compared to a control in an asymptomatic subject may indicate that the subject has PrePF and/or that the subject is at risk for developing IPF.

In some embodiments of the methods of the disclosure, the subject has a mutation in a nucleic acid or amino acid sequence encoding a gene or gene product that is downregulated in a subject having a fibrotic pulmonary disease of the disclosure. In some embodiments of the methods of the disclosure, the subject has a mutation in a nucleic acid or amino acid sequence encoding Surfactant Protein D (SFTPD), Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), Histone Cluster 1 H1 Family Member C (HIST1H1C), YTH Domain Containing 1 (YTHDC1), Plexin A1 (PLXNA1), Serine Peptidase Inhibitor Kazal Type 6 (SPINK6), LDL Receptor Related Protein Associated Protein 1 (LRPAP1), Secretoglobin Family 3A Member 1 (SCGB3A1), H2A Histone Family Member Z (H2AFZ) or Chromosome 1 Open Reading Frame 162 (Clorf162).

TABLE 2 TARGET_GENE_SYM- ORGAN- B-H Fold BOL ISM p-value q-value Change SFTPD Haman 8.19E−15 9.83E−13 −2.262 GAPDH Human 1.46E−09 3.52E−08 −2.096 HIST1H1C Human 3.68E−18 7.80E−16 −2.011 3.63E−16 5.69E−14 −1.964 YTHDC1 Human 1.19E−11 5.38E−10 −1.699 PLXNA1 Human 1.64E−12 1.02E−10 −1.64 SPINK6 Human 3.68E−07 5.04E−06 −1.635 LRPAP1 Human 2.65E−15 3.53E−13 −1.521 SCGB3A1 Human 3.35E−07 4.61E−06 −1.518 H2AFZ Human 3.91E−14 3.91E−12 −1.501 2.95E−11 1.16E−09 −1.493 C1orf162 Human 1.29E−84 7.52E−04 −1.458

In some embodiments of the methods of the disclosure, the subject having PrePF or at risk of developing IPF has a mutation in a sequence encoding MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7.

In some embodiments of the methods of the disclosure, the subject having PrePF or at risk of developing IPF has a mutation in a sequence encoding Telomerase RNA Component (TERC). In some embodiments the polymorphism is rs6793295 comprising (SEQ ID NO: 1).

(SEQ ID NO: 1) AGAAAGAAGT CATGAAAGTA GGAACCACAT TTTTACTCAT CTTTCTGTCT CCAGCAAGCA GCTTACTGCT TTTCATACAC ATTTTGCTTT TATTACTCAT GATTTCAAAG GTGTAATGGT TCAGCCACAT CAATGTAACA AACAGTTCAC ACTGGGCTCT TATAGTCTGG CCTTTAAAAC CTTCACTATT TATGCTTTCA TCTTAACTAC TTTGACCCTC ACAGGTTTAC TCACTAAGAA CTTGAGTTTC AAGAGAAAAG ATGACATGTT TGCTGCTTAA ACAAGCAATA TCTAAAAGCA TATTTAGTTA TAAACGTCTT ACCAAGAATT GATATAATTT TCATTTAAAC ATTTTTATAA ATAGTAGTTT ACAAGATATA GTAAGTACAT CTCTAAAAAT ACAGTGTATT CATGTACCTT GACATAAACT TGTAGTAGTA CCTTAGTTTT ATTCATGTTG TTATATTAAC TACCATCACT TTGAATACAT ACCTGTTCAC B GTACAGTATA GGTCGGTTTA GGTTTATTGC CTTAATTGCT TGGTTTTGAG TTAGTACTGT AGCAAATGCT ATCACACTTT GCATTCCCTA AAAACAGGTA AATTCATTAA GGAAACAGAC AAAGTATATA ATAATCTCGC TACATAAATA TTTCAAGATC AGCTATCTGC ATTCTGATAA AATTGTTTTT AAAATTTAAG CATTCCTTGG ACTTTGAATT GTAAGTTGAT CAAATTCAAA AATGAATTGT TACTGTATTC TTCTCTCCTG GCCCTAAAAT CTATCTAAAA CATGGCATGG GGAGTTTCTT AATGTTTCAG TGTCCATTTC CTGGGTGTTT CCCTCTAGGT TTTTTTTCCT CACCCCTCAA GCTTCTATGT GGATCCCAGC TAGAGCTCAT ACTACTTATC CAACACACAT CATTGTGCAA GCACTCTTTT ATATTCATAC TAGTACTTTT AAGTGTGTGT GCGGTGGGAA AAGGTTACCA ATCACATTTT

In some embodiments of the methods of the disclosure, the subject having PrePF or at risk of developing IPF has a mutation in a sequence encoding Family with sequence similarity 13 member A (FAM13A). In some embodiments the polymorphism is rs2609255 comprising (SEQ ID NO: 2).

(SEQ ID NO: 2) GTATTCATCA ACTCCTATTT CATTCCCTCT TCCTGTGCTC  ACTGGAAGAT GACATTTCCC AGACTTCCAA GAATGTTACT GAGTTCTGGA ATGTAAGTAG AAGGGATAAG TATCACTTCT GTGCTGTGGC GGTTATGGAC CTGTGAACTT TGCACACGCC  TTCTATCTTC TTTTTCAGTG TCCATTTCAG AGGGCATGTT TTCAGATGAA ACCAGTAGAA GATGGAAGCA GCCTGTGACT AGAATCACTG CTTAGGGTCT TGCTGCCTAG GAATCCCACT  CTACCTGCAA CAGACTGTGA AAGAACCGAG AAATACACTG ATTTTGAACA TAGCCCATAC TATAATGGGG ATGTTTGTTA CAGCAGTTAG CATTAAAAAC CTTGGCTAGG CATTGGTCAT  AATTGTAGAA CACAGCAAAT GAAGGGAAAC TGGAACATAG AGGCCAGTGA GAACTTTAGG GTTAATGAAA AATGAGGGCA ACCAGGATAA TTTGGTTCTT K          GCCAAATAGG AAGGTGAAAC CAAAGGTAGA CTGGAGGTCA GAAAATCAGT  CCAGCACATG TGATGTTTTC ATTTAGTTGC CTGTATGTCT GTCTGGTCTC CAGCTCAGCC TGGCTCCTTG AGGTAAGAGG CAGTGGCTGT TCACCTTTGC ATCCCAGCAC CTGGCATACA  ATAGATGGGA TGAAATGTTC AAACTGAGCC TAAGCTTCAG GGTGCTTATC AAAGCAGGGA AGATACACAA GAGGAGATGA TTCAGGTCCA GGGCAGGTCA GGTATCTAAA CCCAGTCTCT  TAGGAAGCTG GATCCTCCGA ACCAGGGAGA ACAAGCTGGA TATGCACTGG ATTTCCCAGC AGTACTGATC TAGAGACTCT CATAGAGTCC CTTTTATTCC TTGGCCTAGG GTTACAACTG  CTTATAGCAT CTGGAAAGAC TCAACACCTC AAAAGAGACT TTCAGTAGAT ACAGCAAATA CACTCATGGA ATTGATAATT AAGCTTCAAT

In some embodiments of the methods of the disclosure, the subject having PrePF or at risk of developing IPF has a mutation in a sequence encoding Telomerase Reverse Transcriptase (TERT). In some embodiments the polymorphism is rs2736100 comprising (SEQ ID NO: 3).

(SEQ ID NO: 3) ATTGTCGTTG TTTGCTTTTG TTTATTGAGA CAGTCTCACT  CTGTCACCCA GGCTGGAGTG TAATGGCACA ATCTCGGCTC ACTGCAACCT CTGCCTCCTC GGTTCAAGCA GTTCTCATTC CTCAACCTCA TGAGTAGCTG GGATTACAGG CGCCCACCAC  CACGCCTGGC TAATTTTTGT ATTTTTAGTA GAGATAGGCT TTCACCATGT TGGCCAGGCT GGTCTCAAAC TCCTGACCTC AAGTGATCTG CCCGCCTTGG CCTCCCACAG TGCTGGGATT  ACAGGTGCAA GCCACCGTGC CCGGCATACC TTGATCTTTT AAAATGAAGT CTGAAACATT GCTACCCTTG TCCTGAGCAA TAAGACCCTT AGTGTATTTT AGCTCTGGCC ACCCCCCAGC  CTGTGTGCTG TTTTCCCTGC TGACTTAGTT CTATCTCAGG CATCTTGACA CCCCCACAAG CTAAGCATTA TTAATATTGT TTTCCGTGTT GAGTGTTTCT K          TAGCTTTGCC CCCGCCCTGC TTTTCCTCCT TTGTTCCCCG TCTGTCTTCT  GTCTCAGGCC CGCCGTCTGG GGTCCCCTTC CTTGTCCTTT GCGTGGTTCT TCTGTCTTGT TATTGCTGGT AAACCCCAGC  TTTACCTGTG CTGGCCTCCA TGGCATCTAG CGACGTCCGG  GGACCTCTGC TTATGATGCA CAGATGAAGA TGTGGAGACT CACGAGGAGG GCGGTCATCT TGGCCCGTGA GTGTCTGGAG  CACCACGTGG CCAGCGTTCC TTAGCCAGTG AGTGACAGCA  ACGTCCGCTC GGCCTGGGTT CAGCCTGGAA AACCCCAGGC ATGTCGGGGT CTGGTGGCTC CGCGGTGTCG AGTTTGAAAT  CGCGCAAACC TGCGGTGTGG CGCCAGCTCT GACGGTGCTG  CCTGGCGGGG GAGTGTCTGC TTCCTCCCTT CTGCTTGGGA ACCAGGACAA AGGATGAGGC TCCGAGCCGT TGTCGCCCAA  CAGGAGCATG

In some embodiments of the methods of the disclosure, the subject having PrePF or at risk of developing IPF has a mutation in a sequence encoding Desmoplakin (DSP). In some embodiments the polymorphism is rs2076295 comprising (SEQ ID NO: 4).

(SEQ ID NO: 4) ATTTGGGAAC CTTTAAAAAA TATTCTGGCT TCAAAAATAC  TCCATATTTA CATCTTTGGT TCTATCTGAA GTAAAGCCGT GATGGTGTGC GTAAGTGAAA CAGGTGCAAA GGGGCAACAA CAAAGGGCGC CTCTCTTTGT CTTTGTGTCG CAGGCGGAGA  TGGACATGGT GGCCTGGGGT GTGGACCTGG CCTCAGTGGA GCAGCACATT AACAGCCACC GGGGCATCCA CAACTCCATC GGCGACTATC GCTGGCAGCT GGACAAAATC AAAGCCGACC  TGGTACTTGT CTGTGTTTCA TTTTAGAGTC TTCAAAATAT CTACCGAAGG ATCGTGTAAT TACTCAATCC CAGGGAGTTT CTTCTGAAAC ATTGCTATTA TTTCTTTCCC AGAAGACTGG  AAATGTTTAG AAATCCCACT TCTTAAATGG GGAAGTGGAA TCAGTAGCCC TATTAGAGAT TATGTTAACA CTTGAAGAGG AGTTAAACCA GAGGCTGAGG K          TGTGCAAACA CTCATTTGCA GTTTGTGAAT AAGTCTCTTT AGGGGTGGCA  GTTTGTTTCT GCGGTAAGCA GAACATCTTT TTGAATAGGG GAAATGCAAC AGTCTTATAC AGTAGTTTGT GTCATTGGTG  AATCCTTTCC TAGGTGGTAA TTAAAACATT ATTTCTACTG  AGCAAAGCCA TATGTCATCC CGACACCCGC TCCCATGCTG AAAAAAGTCA GACTTGAAAC TGGGTTGAGA ATTACAGCAT  AAAATCATAA CTGATCTTAA GTGCTTAGTT TCCCGCAGGT  CTCTACACTT GTAAATCACT AAACTTTTTT TTTTTTTTTT TACCTGAGAC CATAGCTTCT CATCCTCATT TCTTCTTCTG  GCTTTTTGGG GCTTACTTTT GTCCACCTGA GCCCCTGACC  AACTTTCTCC TTCATTTCTC TAAGACCTAG GGAATCCTAA  ATGATGTCTT TAAACTTTAA GACAATTTTC TAACACGTGA  GTCTTTAAGT

In some embodiments of the methods of the disclosure, the subject having PrePF or at risk of developing IPF has a mutation in a sequence encoding Zinc-alpha 2-Glycoprotein 1 (AZGP1). In some embodiments the polymorphism is rs4727443 comprising (SEQ ID NO: 5).

(SEQ ID NO: 5) CCCAACCCAA ATAAGCACTA TAACCTCTTG TTATTCACTT  CTCATGCAAC CAGTCTTCTG TTCTCTGTGA GTCTTTAGGA AATGAGGAGC ATGATCTTCT AGCAGTAAAA CACCTGTAGA GAATTGCCTT ATGTTTTTTG TTTGTTTATT TGTTTGTGTG  CTTTGGTTTG GTTTGCTTTT TTTTTTTTTT TTTTTTTTTT TTTGAGATGG AGTCTCGCCC TGTTGCCCAG GCTGGAGTGT AGTGGCGAAA TCTCGGCTCA CTGCAACCTC CACCTCCCTG  GTTCAAGCAA TTCCCCTGTC TCAGCCTCCC GAGTAGCTGA GATTACAGGT GCACACCACC ACGCCCGGCT AATTTTTTTG TATTTTTAGT AGAGATGGGG TTTCACCATG TTGGCCAGAC  TGGTCTCGAA CTTCTGACCT CAGGCAATCC GCCTGCCTCA GCCTCCCAAA GCGCTGGGAT TACAGGCATG AGCCACTGCG CCCCGCCTCC ATGTTAATCA M          TCTTTCTGAT TTCAAATAAC TCATTATCCC CATGACCTTA TGGATTTGTT  TTTCCTCTTC ATCCACAAAA TTCTCCAGAG AAGTCTCCCT TGTTATCTCT TGGCTGTGCT TTCTATCTCA CCAGTTATCT  TTCTCCAAAG AGCTTCCTCT GCAAAGAAGC TTTGTATATG  AAGACCATGT GGGGGCTGAA TCAAGACCAA GTTTCACAAC CTAAAAGTAG TTCACAAAGC TTCCTTGCCT CTATTCTCTG  CAAATCTGTA AACTCTTCAG CTGACCCAAT TTCTCTCTTT  AGCCTTCAGA GATTATTTTA TTTTATTTTA TTTCATTTCA TTTCATTTCA TTTTGACAGA ATCTAGCTCT GTCGCCCAGG  CTGGAGTGCA GTGGCACCAT CTTTGCTCAC TGCAACCTCC  CCCTCACAGG TTCAAGCAAC TGTCCTGCCT CAGCCTCCCG AGTAGCTGGG ATTACAGGCG TGAGCCACCA CGCCCAGCTG ATTTTTTTTT 

In some embodiments of the methods of the disclosure, the subject having PrePF or at risk of developing IPF has a mutation in a sequence encoding Oligonucleotide/oligosaccharide-binding Fold Containing 1 (OBFC1). In some embodiments the polymorphism is rs11191865 comprising (SEQ ID NO: 6).

(SEQ ID NO: 6) CCTCTACTGC CGTACACCCC ACCACTCAGC CTTGGAGTGC  CTGTGTGCAG AGCAGGGCTG AGGCATGGTG CTGCTTTGGT GGTCTAGGTT TGCTGCAGGG CCAGGTGGCC TGAGCTCCAG GCAGGATCTC TGGCTGCACT CAGCCCTTTC TGCCTCCCCA  AATGCTCTAT ATCACTATTT GTACACTGAG CAGAGTAAAG TTAGAGAGAA CTGTTTTATA GAATAGGGCT GGCCCCCGCT CCCCTGGCCT ACGTGATGGT CCTTCCTGGC TGCCAGGTAC  TTGTTTGTAT TAGAGACAGA CACTCCACAG GGTCTGTTGT GGCCCACAGC ACATAGGCAA TCAGAGGCAG AAAGCAGAGC TGTTTGGACC CACAGAGGGC CGGCTGTCTG CCACTGAAAT  GTCTTTCCAG TTGGTTGAGA AGCAGCAGGA TGCTCTGCTG GTGATGTCTG AAAGTCCCAG GATTCTTTGG GTCTCCAAGG AGATCCTAGC ATATACCACT R          TCGTGGTTTT  AATAAAGAGC AAAAACACTT TCAGATGGGG AGAAGAGTGG  AACAAAAGGT ATTCTTCCTG GGTTGAAGTC TGGGGGAAAG GCATTGAGAA GACTGGGCTA ATGGCACAAA CCAATGAAGT  ACTCAAGTCA CCTGTGATGG AGGCCAGTCA TCCAATGGTA  TCAACTTTGT ATGTGGCAAC ACTTAATAAA AATCTGAACA GGTCTTCACT TGTGGACACA GTAGACTTTC TTGAAAAAGG  ACAGAAAAGT GAGCCCTGTG AATTTTCATC TCACGGACTG  ACAACAATGA CTTGCCTTTA AGGACAGTCA CTCAAGATGA AGATGCAACA AAACCCTTCC AGTTCCAAGT GGCTGATGAA  AAAAAAAAAA TCTTAAAAGC ATCACAGAAC AACGGAGAAA  GAGATCAGAA GACTATAACA GATAGTTTGA ATTTTAAAAC TCAGAGAAAA GCAACTGAGG AGGAAATACA CTGCTTAGAA  AGAAGAAACT

In some embodiments of the methods of the disclosure, the subject having PrePF or at risk of developing IPF has a mutation in a sequence encoding Mucin 5B (MUC5B). In some embodiments the polymorphism is rs35705950 comprising (SEQ ID NO: 7).

SEQ ID NO: 7) TGGACGGCCT CTGAAGGGGT CTGTGGGGTC CTGGACGGGT  CCCCATTCAT GGCAGGATTA ACCCCCCTCG GGTTCTGTGT GGTCCAGGCC GCCCCTTTGT CTCCACTGCC CCCTGGCCAG AATGAGGGAC AGTGACCCAC CCAGGGCTGG GCCTGGCTCA  GACTCCGTCA GAGCCGCAGG GCAAGTTCCT GGCACGTCCG AGGTGGGAGG CTCCTCTGCG CTCCAGGAGG CTGTGCCTGG CCCCCCTTCC CGGCAGGAAC CGGCTGTGTC CCTTTCCTTC  CTTTATCTTC TGTTTTCAGC D          CCTTCAACTG  TGAAGAGGTG AACTCTTCAA ACACGCTGAG CAAACAGGCC  CGACTCCCAG GGCCGCATCC GGGATGTCTC AATAGCTGTG GCCTTGACGT CCACCTCGGA CCCCTGCCCC GGACCCAGCC  CAGTTCCCAA TGGGCCCTCT GCCCGGGGAG GTGCCTAGTG  GGAGGGACGA GGGCAAAGTC GGGGCCCCCA CTTGTTTGGT GTCACTGTGT GCCAGCGGCC ACTGGCGGGC GAGGCTGTTC  CAGGGTGGAG GCGGGGAGGG TTGGACCACA GGCACTGAGC  GGGGACAGAG

In some embodiments of the methods of the disclosure, the subject having PrePF or at risk of developing IPF has a mutation in a sequence encoding ATPase Phospholipid Transporting 11A (ATP11A). In some embodiments the polymorphism is rs12787690 comprising (SEQ ID NO: 8).

(SEQ ID NO: 8) GTCATTGGTC AAATGTGGCC TGTATCTAAA TTCCAACTGT  TAGAATCATA GACATCTAGA GCTTACGTCA GTTTTAGATA  TTTCTTATGA ATTCTCAGAA TTCATAGATT CTCATTTTTA TTCTTAGACT TCTCAGATAT TCCGTTTTTG ATAGTATACC  CTTCTGAGTC TAATATGTCC TAAAGTGCGA ACTTGTACAA TTTttttttt tttttttttt tttttttttt t K          tgataaggag ttttactctg tcacccaggc  tggagtgcag tgacccgatc tcggctcact gcaacctctg  cctcccgggt tcaagtgatt gtgatgtctc agtctcccaa  gtagctggga ttacaggctc ctgccaccac atgcctagct  aattgttata ctttagtaga aatggggctt cgccgtgtta  gtcaggctgg tcttgtactc ctgacctcag ttgatctgcc  taccttggcc cccaaggtgc tgggattaca ggcatgagcc accgcgcctg accCAGCTTC TTAAATTATT CTGGGCCACC  AGTAATGTGA ATCATGtaaa ttaaaatata taattaaaCA  AAATCATATA GCGATTAGAG ATAATAGTTG TGAAATGCTT GAAAAATCAT AGGCATTTAA TAAATAGAAG CCATTCCAAT  TAGGATTCTT CTTGATTTTT TTTCAAGACC AAAAAAATAC  TCttttaaat atttattata ataCTCCATG

In some embodiments of the methods of the disclosure, the subject having PrePF or at risk of developing IPF has a mutation in a sequence encoding Isovaleryl-CoA dehydrogenase (IVD)/Dispatched RND Transporter Family Member 2 (DISP2). In some embodiments the polymorphism is rs2034650 comprising (SEQ ID NO: 9).

(SEQ ID NO: 9) aggctgcagt tagtcatgac tgcgcgctgc actccagcct  gggtgacaaa gtgaggccct gtctcaaaaa caataaaaaa TTTAAAAGAG CTGAGCATGG AGGCcacttt gggaggctga ggcaggcaga tctcttaagc ccaggagtct gagaccagcc  tgggcgacat gatgaagccc catctctaca aaaaatacaa aaaaattagc tgagctttat ggcaaatccc tgtaatccca gttacctagg aggcccaggc aggaagatgg cttgagccca  aaaggttgag gctgtagtga gctgtgatca tgaacagagt gagaccctgt ttcaaaacaa aatgaaaaac aaacaaacaa aaaaaCCAAG AAAACAAGAA AACAAAAACT ATACAATGAT  GAGCCAAAAA GCAAGATATG GAAGAatata tatatatata  tatatatata tataGTATGA GTCCAGCTAT AGAAAGTTTG AAATCAGGCA ACCTAAACAA TATTGTTCAG GGATCTATAC AGAGGCAGGA AGCCATTGAG AAAGGTAAGG GGAGGATTAT  CACCAAATTC AGGATGGTGG CTCCCCTGGG GAGAATATGT CAAGGAGGGG CACATGGGCT TGGAATACTG TCTTCATTGA  CCTGCGTGTT GGGTACACAG GAGTTTGTTA TTTTTCACAC  TGCATATGTG CATGTATATA CTCTCCCATA TATACCATGC ATTTCACACA AGAACACAAA GGCTGTGTGG CTCTGCTCTG  CCCCTTTCCC CTTCCAGCTC CCATTCTCGT C Y          TCAGCTAGCA GAGGAGGGTC AGGGTCTTTT  AGCACAGCTT CCTTCTGTCT CTGAGTGGGT CAGAGGAGTA  CGGGGATGAG GGCCTCCCTT CTGCGGCTGG GCTCTGGCCA  CTCCAGGGTG GGAAGGCCTG GAGAAAACAG GGCCAGGCAA  AGCCGGCTGG CCCTGCTGTT TCTGCCAATG CTGGGATTAG  GCCAGGGCTC TGGCCCACCT GTCATTTCAC TCATTCAGCA  TGAACATAGC CACTGAGCAC TTACTGTGAG CCCCGGGTGC  TATTGGGAGA GTTCAGATAA GTGAGAGAGG GTCTTTGACC  TCAAAGATCT TACAGAGAGG ACCGTATACA CAAATAACAG  TATACCAGCA AAATGTGAGC TAAGTGTCAT GTGACTACTC atctactctt tcaataaata tttgttgtgc acctattaca  tgccaggaac tgtgctggat ggtgatcatg taaagacagt  caaatcacag tcctagctct cagattcaca gcctgcctaa tgctggggaa acTGGAAT

In some embodiments of the methods of the disclosure, the subject having PrePF or at risk of developing IPF has a mutation in a sequence encoding Dipeptidyl Peptidase 9 (DPP9). In some embodiments the polymorphism is rs12610495 comprising (SEQ ID NO: 10).

(SEQ ID NO: 10) CCAGCCAGAA GGGGCGCAGT TTGTTAGTTC AGCTCCTCCT  GAGACAGAAA TAAAGACACG AACCAAAGGA CATCAGCACT  TACAGGGCTC TCAGGTCACA CACAGGATGT CCGCGCCCAC TGCAGAGCTG CAGGTCCCCT CCAGGGCAGT GGGGAGCCAC  AAGCAGCGTT AGGCAGCGGC TGGGACCAGG ACCGCCTGAG CACTCAAGAA CCCCCACTGC CCCAAGCACT GCTGGCAGCA AGCCCAGAAA ACTGAGCCCG GGGAGCTCCT CTGAGCGGCC TAAGCACCCC TCTAAGCTGT GCTGCCCCAA TTCAAGCCTG GCTCACGGCA GCAAAGAAAA AATGTGACCT TCGGAGCTCC CAAAGGGGCC ACCCATAAGC TGAGAGCCTG CCCGGAAGCA  CTTATAGACC CGCGTGGCTT GTTTTCATTG CAAAGAACAA  TAAAAATTAT CTTGCCTCTG ATCACCACTG ATAGCCCAAG AAGCAAAAAT TCGATCCCGG D          GATGAGAAAT  GAAATGAAAC ATCGCGAGAA ACTTCCAGGA ATCTTCTGGA  TGTGGCTAGA CTCTTTAGCT TGAGCTTCCA GACAGGCCGA  GGCTTGGTGC TGGAGCCTGG CCCTCCGCTG ACCTCTCTTC  TACCCGGGGG CACAGCCCGG ATTGCAGAGA GGCTGGCGCA  AGAGTGAGGG AGCGAGGGCT AGCCTGTGAT GGGCTTTCTC  CACCTAGCAC CACCCTATGC TGTGGCTCAG GGGAGTCAAG  AGTTTACACA GCTGCAGAGA TGGATTCCAG GCCACTTACT  CAAGTCTACC TACTCCTTCC TTCGGCCAAT CAGCTGGGTG  CCTCTGCGGC CTGTGACACC ACCAGCAAAC AGCTCCAGAC  CTCCTAGCAT GGTCTCTGTC AAGGCTGGGT GGCAGATCTG  TGATCTCCTT TTTAAATTTT TCATTTTTTT TAAGAGATGG  GGTCTTGCTA TATTGCCCAG GCTGGTCTCA AACTCCTGGG  CTCCAGCGAT

In some embodiments of the methods of the disclosure, the wild type human MUC5B gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_002458.2):

(SEQ ID NO: 11)     1 cacccggccc ggctccctcc ctgcccgtcc ccgtcccccc acccgtgcca gcccccagga    61 tgggtgcccc gagcgcgtgc cggacgctgg tgttggctct ggcggccatg ctcgtggtgc   121 cgcaggcaga gacccagggc cctgtggagc cgagctggga gaatgcaggg cacaccatgg   181 atggcggtgc cccgacgtcc tcgcccaccc ggcgcgtgag ctttgttcca cccgtcactg   241 tcttccccag cctgagcccc ctgaacccgg cgcacaatgg gcgggtgtgc agcacctggg   301 gtgacttcca ctacaagacc ttcgacggcg acgtcttccg cttccctggc ctttgcaact   361 acgtgttctc tgagcactgc cgcgccgcct acgaggactt caacgtccag ctacgccgag   421 gcctagtggg ctccaggcct gtggtcaccc gtgttgtcat caaggcccag gggctggtgc   481 tggaggcgtc caacggctcc gtcctcatca atgggcagcg ggaggagctg ccttacagcc   541 gcactggcct cctggtggag cagagcgggg actacatcaa ggtcagcatc cggctggtgc   601 tgacattcct gtggaacgga gaggacagtg ccctgctgga gctggatccc aaatacgcca   661 accagacctg tggcctgtgt ggggacttca acggcctccc ggccttcaac gagttctatg   721 cccacaacgc caggctgacc ccgctccagt ttgggaacct gcagaagttg gatgggccca   781 cggagcagtg cccggacccg ctgcccttgc cggccggcaa ctgcacggac gaggagggca   841 tctgccaccg caccctgctg gggccggcct ttgcggagtg ccacgcactg gtggacagca   901 ctgcgtacct ggccgcctgc gcccaggacc tgtgccgctg ccccacctgc ccgtgtgcca   961 cctttgtgga atactcacgc cagtgcgccc acgcgggggg ccagccgcgg aactggaggt  1021 gccctgagct ctgcccccgg acctgccccc tcaacatgca gcaccaggag tgtggctcac  1081 cctgcacgga cacctgctcc aacccccagc gcgcgcagct ctgcgaggac cactgtgtgg  1141 acggctgctt ctgcccccca ggcacggtgc tggatgacat cacgcactct ggctgcctgc  1201 ccctcgggca gtgcccctgc acccacggcg gccgcaccta cagcccgggc acctccttca  1261 acaccacctg cagctcctgc acctgctccg gggggctatg gcagtgccag gacctgccgt  1321 gccctggcac ctgctctgtg cagggcgggg cccacatctc cacctatgat gagaaactct  1381 acgacctgca tggtgactgc agctacgttc tgtccaagaa atgtgccgac agcagcttca  1441 ccgtgctggc tgagctgcgg aagtgcggcc tgacggacaa cgagaactgc ctgaaagcgg  1501 tgacgctcag cctggacggc ggggacacgg ccatccgggt ccaagcggac ggcggcgtgt  1561 tcctcaactc catctacacg cagctgcccc tgtcggcagc caacatcacc ctgttcacac  1621 cctcgagctt cttcatcgtg gtgcagacag gcctggggct gcagctgctg gtgcagctgg  1681 tgccactcat gcaggtgttt gtcaggctgg accccgccca ccagggccag atgtgcggcc  1741 tgtgtgggaa cttcaaccag aaccaggctg acgacttcac ggccctcagc ggggtggtgg  1801 aggccacggg cgcagccttc gccaacacct ggaaggccca ggctgcctgt gccaatgcca  1861 ggaacagctt tgaggacccc tgctccctca gtgtggagaa tgagaactac gcccggcact  1921 ggtgctcgcg cctgaccgat cccaacagtg ccttctcgcg ctgccactcc atcatcaacc  1981 ccaagccctt ccactcgaac tgcatgtttg acacctgcaa ctgtgagcgg agcgaggact  2041 gcctgtgcgc cgcgctgtcc tcctatgtgc acgcctgtgc cgccaagggc gtacagctca  2101 gcgactggag ggacggcgtc tgcaccaagt acatgcagaa ctgccccaag tcccagcgct  2161 acgcctacgt ggtggatgcc tgccagccca cttgccgcgg cctgagtgag gccgacgtca  2221 cctgcagcgt ttccttcgtg cctgtggacg gctgcacctg ccccgcgggc accttcctca  2281 atgacgcggg cgcctgtgtg cccgcccagg agtgcccctg ctacgctcac ggcaccgtgc  2341 tggctcctgg agaggtggtg cacgacgagg gcgccgtgtg ttcatgtacg ggtgggaagc  2401 taagctgcct gggagcctct ctgcagaaaa gcacagggtg tgcagccccc atggtgtacc  2461 tggactgcag caacagctcg gcgggcaccc ctggggccga gtgcctccgg agctgccaca  2521 cgctggacgt gggctgtttc agcacacact gcgtgtccgg ctgtgtctgt cccccggggc  2581 tggtgtcgga tgggagtggg ggctgcattg ccgaggagga ctgcccctgt gtgcacaacg  2641 aggccaccta caagcctgga gagaccatca gggtcgactg caacacctgc acctgcagga  2701 accggaggtg ggagtgcagc caccggctct gcctgggcac ctgcgtggcc tacggggatg  2761 gccacttcat cacctttgat ggcgatcgct acagctttga aggcagctgc gagtacatct  2821 tggcccagga ctactgtggg gacaacacca cccacgggac cttccgcatc gtcaccgaga  2881 acatcccctg tgggaccacc ggcaccacct gctccaaggc catcaagctc ttcgtggaga  2941 gctacgagct gatcctccaa gaggggacct ttaaggcggt ggcgagaggg ccgggtgggg  3001 acccacccta caagatacgc tacatgggga tcttcctggt catcgagacc cacgggatgg  3061 ccgtgtcctg ggaccggaag accagcgtgt tcatccgact gcaccaggac tacaagggca  3121 gggtctgcgg cctgtgcggg aacttcgacg acaatgccat caatgacttt gccacgcgta  3181 gccggtccgt ggtgggggac gcactggagt ttgggaacag ctggaagctc tccccctcct  3241 gcccggacgc cctggcaccc aaggacccct gcacggccaa ccccttccgc aagtcctggg  3301 cccagaagca gtgcagcatc ctccacggcc ccaccttcgc cgcctgccgc tcccaggttg  3361 actccaccaa gtactacgag gcctgcgtga acgacgcgtg tgcctgcgac tcgggtggcg  3421 actgcgagtg tttctgcacg gctgtggctg cctacgccca ggcctgccac gacgcgggcc  3481 tgtgtgtgtc ctggcggact ccggacacct gccccttgtt ctgtgacttc tacaacccac  3541 atgggggctg tgagtggcac taccagccct gcggggcacc ctgcctaaaa acctgccgga  3601 accccagtgg gcactgcctg gtggacctgc ctggcctgga aggctgctac ccgaagtgcc  3661 cacccagcca gcccttcttc aatgaggacc agatgaagtg cgtggcccag tgtggctgct  3721 acgacaagga cggaaactac tatgacgtcg gtgcaagggt ccccacagcg gagaactgcc  3781 agagctgtaa ctgcacaccc agtggcatcc agtgcgctca cagccttgag gcctgcacct  3841 gcacctatga ggacaggacc tacagctacc aggacgtcat ctacaacacc accgatgggc  3901 ttggcgcctg cttgatcgcc atctgcggaa gcaacggcac catcatcagg aaggctgtgg  3961 catgtcctgg aactccagcc acaacgccat tcaccttcac caccgcctgg gtcccccact  4021 ccacgacaag cccggccctc ccggtctcca ccgtgtgtgt ccgcgaggtc tgccgctggt  4081 ccagctggta caatgggcac cgcccagagc ccggcctggg aggcggagac tttgagacgt  4141 ttgaaaacct gaggcagaga gggtaccagg tatgccctgt gctggctgac atcgagtgcc  4201 gggcggcgca gcttcccgac atgccgctgg aggagctggg ccagcaggtg gactgtgacc  4261 gcatgcgggg gctgatgtgc gccaacagcc aacagagtcc cccgctctgt cacgactacg  4321 agctgcgggt tctctgctgc gaatacgtgc cctgtggccc ctccccggcc ccaggcacca  4381 gccctcagcc ctccctcagt gccagcacgg agcctgctgt gcctacccca acccagacca  4441 cagcaaccga aaagaccacc ctatgggtga ccccgagcat ccggtcgacg gcggccctca  4501 cctcgcagac tgggtccagc tcaggccccg tgacggtcac cccctcggcc ccaggtacca  4561 ccacctgcca gccccggtgt cagtggacag agtggtttga tgaggactac cccaagtctg  4621 aacaacttgg aggggacgtt gagtcctacg ataagatcag ggccgctgga gggcacttat  4681 gccagcagcc taaggacata gagtgccagg ccgagagctt ccccaactgg accctggcac  4741 aggtggggca gaaggtgcac tgtgacgtcc acttcggcct ggtgtgcagg aactgggagc  4801 aggagggcgt cttcaagatg tgctacaact acaggatccg ggtcctctgc tgcagtgacg  4861 accactgcag gggacgtgcc acaaccccgc caccgaccac agagctggag acggccacca  4921 ccaccaccac ccaggccctg ttctcaacgc cgcagcctac gagtagcccg gggctgacca  4981 gggctccccc ggccagcacc acagcagtcc ccaccctctc agaaggactg acatccccca  5041 gatacacaag cacccttggt acagccacca cgggaggccc cacgacgcct gcaggctcca  5101 cagaacccac tgtcccaggg gtggccacat ccacccttcc aacacgctca gcccttccag  5161 ggacgacggg gagcttgggc acatggcgcc cctcacagcc acccacgctg gccccaacaa  5221 caatggcaac ctccagagct cgcccgacag gcacagccag caccgcttcc aaagagccgc  5281 tgaccacgag cctggcgcca acactcacga gcgagctgtc cacctctcag gccgagacca  5341 gcacgcccag gacagagacg acaatgagcc ccttgactaa caccaccacc agccagggca  5401 cgacccgctg tcaaccgaag tgtgagtgga cagagtggtt tgacgtggac ttcccaacct  5461 caggggttgc aggcggggac atggaaactt ttgaaaacat cagggctgct gggggcaaga  5521 tgtgctgggc accaaagagc atagagtgcc gggcggagaa ctaccccgag gtaagcatcg  5581 accaggtcgg gcaggtgctg acctgcagcc tggagacggg gctgacctgc aagaacgaag  5641 accagacagg caggttcaac atgtgcttca actacaacgt gcgtgtgctt tgctgtgacg  5701 actacagcca ctgccccagt accccagcca ccagctccac ggccacgccc tcctcaactc  5761 cggggacgac ctggatcctc acaaagccga ccacaacagc cactacgact gcgtccactg  5821 gatccacggc caccccgacc tccaccctga gaacagctcc ccctcccaaa gtgctgacca  5881 ccacggccac cacacccaca gtcaccagct ccaaagccac tccctcctcc agtccaggga  5941 ctgcaaccgc ccttccagca ctgagaagca cagccaccac acccacagct accagcgtta  6001 cacccatccc ctcttcctcc ctgggcacca cctggacccg cctatcacag accaccacac  6061 ccacggccac catgtccaca gccacaccct cctccactcc agagactgcc cacacctcca  6121 cagtgcttac cgccacggcc accacaactg gggccaccgg ctctgtggcc accccctcct  6181 ccaccccagg aacagctcac actaccaaag tgccaactac cacaaccacg ggcttcacag  6241 ccaccccctc ctccagccca gggacggcac tcacgcctcc agtgtggatc agcacaacca  6301 ccacacccac aaccagaggc tccacggtga ccccctcctc catcccgggg accacccaca  6361 ccgccacagt gctgaccacc accaccacaa ctgtggccac tggttctatg gcaacaccct  6421 cctctagcac acagaccagt ggtactcccc catcactgac caccacggcc actacgatca  6481 cggccaccgg ctccaccacc aacccctcct caactcctgg gacaactccc atccccccag  6541 tgctgaccac caccgccacc acacctgcag ccaccagcaa cacagtgact ccctcctctg  6601 ccctagggac cacccacaca cccccagtgc cgaacaccat ggccaccaca cacgggcgat  6661 ccctgccccc cagcagtccc cacacggtgc gcacagcctg gacttcggcc acctcgggca  6721 tcttgggcac cacccacatc acagagcctt ccacggtgac ttcccacacc ctagcagcaa  6781 ccaccggtac cacccagcac tcgactccag ccctttccag ccctcaccct agcagcagaa  6841 ccaccgagtc acccccttct ccagggacga ccaccccggg ccacaccacg gccacctcca  6901 ggaccacagc cacggccaca cccagcaaga cccgcacctc gaccctgctg cccagcagcc  6961 ccacatcggc ccccataacc acggtggtga ccatgggctg tgagccccag tgtgcctggt  7021 cagagtggct ggactacagc taccccatgc cggggccctc tggcggggac tttgacacct  7081 actccaacat ccgtgcggcc ggaggggccg tctgtgagca gcccctgggc ctcgagtgcc  7141 gtgcccaggc ccagcctggt gtccccctgc gggagttggg ccaggtcgtg gaatgcagcc  7201 tggactttgg cctggtctgc aggaaccgtg agcaggtggg gaagttcaag atgtgcttca  7261 actatgaaat ccgtgtgttc tgctgcaact acggccactg ccccagcacc ccggccacca  7321 gctctacggc catgccctcc tccactccgg ggacgacctg gatcctcaca gagctgacca  7381 caacagccac tacgactgag tccactggat ccacggccac cccgtcctcc accccaggga  7441 ccacctggat cctcacagag ccgagcacta cagccaccgt gacggtgccc accggatcca  7501 cggccaccgc ctcctccacc caggcaactg ctggcacccc acatgtgagc accacggcca  7561 cgacacccac agtcaccagc tccaaagcca ctcccttctc cagtccaggg actgcaaccg  7621 cccttccagc actgagaagc acagccacca cacccacagc taccagcttt acagccatcc  7681 cctcctcctc cctgggcacc acctggaccc gcctatcaca gaccaccaca cccacggcca  7741 ccatgtccac agccacaccc tcctccactc cagagactgt ccacacctcc acagtgctta  7801 ccaccacggc caccacaacc ggggccaccg gctctgtggc caccccctcc tccaccccag  7861 gaacagctca cactaccaaa gtgctgacta ccacaaccac gggcttcaca gccaccccct  7921 cctccagccc agggacggca cgcacgcttc cagtgtggat cagcacaacc accacaccca  7981 caaccagagg ttccacggtg accccctcct ccatcccggg gaccacccac acccccacag  8041 tgctgaccac caccaccaca actgtggcca ctggttctat ggcaacaccc tcctctagca  8101 cacagaccag tggtactccc ccatcactga ccaccacggc cactacgatc acggccaccg  8161 gctccaccac caacccctcc tcaactccag ggacaacacc tatcccccca gtgctgacca  8221 ccaccgccac cacacctgca gccaccagca gcacagtgac tccctcctct gccctaggga  8281 ccacccacac acccccagtg ccgaacacca cggccaccac acacgggcga tccctgtccc  8341 ccagcagtcc ccacacggtg cgcacagcct ggacttcggc cacctcaggc accttgggca  8401 ccacccacat cacagagcct tccacgggga cttcccacac cccagcagca accaccggta  8461 ccacccagca ctcgactcca gccctgtcca gccctcaccc tagcagcagg accaccgagt  8521 cacccccttc tccagggacg accaccccgg gccacaccag ggccacctcc aggaccacgg  8581 ccacggccac acccagcaag acccgcacct cgaccctgct gcccagcagc cccacatcgg  8641 ccccaataac cacggtggtg accatgggct gtgagcccca gtgtgcctgg tcagagtggc  8701 tggactacag ctaccccatg ccggggccct ctggcgggga ctttgacacc tactccaaca  8761 tccgtgcggc cggaggggcc gtctgtgagc agcccctggg cctcgagtgc cgtgcccagg  8821 cccagcctgg tgtccccctg cgggagttgg gccaggtcgt ggaatgcagc ctggactttg  8881 gcctggtctg caggaaccgt gagcaggtgg ggaagttcaa gatgtgcttc aactatgaaa  8941 tccgtgtgtt ctgctgcaac tacggccact gccccagcac cccggccacc agctctacgg  9001 ccacgccctc ctccactcca gggacgacct ggatcctcac agagcagacc acagcagcca  9061 ctacgaccgc aaccactgga tccacggcca tcccgtcctc caccccggga acagctcccc  9121 ctcccaaagt gctgaccagc acggccacca cacccacagc caccagttcc aaagccactt  9181 cctcctccag tccaaggact gcaaccaccc ttccagtgct gacaagcaca gccaccaaat  9241 ccacagctac cagctttaca cccatcccct ccttcaccct tgggaccacc gggaccctcc  9301 cagaacagac caccacaccc atggccacca tgtccacaat ccacccctcc tccactccgg  9361 agaccaccca cacctccaca gtgctgacca cgaaggccac cacgacaagg gccaccagtt  9421 ccatgtccac cccctcctcc actccgggga cgacctggat cctcacagag ctgaccacag  9481 cagccactac aactgcagcc actggcccca cggccacccc gtcctccacc ccagggacca  9541 cctggatcct cacagagccc agcactacag ccaccgtgac ggtgcccacc ggatccacgg  9601 ccaccgcctc ctccacccgg gcaactgctg gcaccctcaa agtgctgacc agcacggcca  9661 ccacacccac agtcatcagc tccagagcca ctccctcctc cagtccaggg actgcaaccg  9721 cccttccagc actgagaagc acagccacca cacccacagc taccagcgtt acagccatcc  9781 cctcttcctc cctgggcacc gcctggaccc gcctatcaca gaccaccaca cccacggcca  9841 ccatgtccac agccacaccc tcctctactc cagagactgt ccacacctcc acagtgctta  9901 ccaccacgac caccacaacc agggccaccg gctctgtggc caccccctcc tccaccccag  9961 gaacagctca cactaccaaa gtgccgacta ccacaaccac gggcttcaca gccaccccct 10021 cctccagccc agggacggca ctcacgcctc cagtgtggat cagcacaacc accacaccca 10081 caaccagagg ctccacggtg accccctcct ccatcccggg gaccacccac accgccacag 10141 tgctgaccac caccaccaca actgtggcca ctggttctat ggcaacaccc tcctctagca 10201 cacagaccag tggtactccc ccatcactga ccaccacggc cactacgatc acagccaccg 10261 gctccaccac caacccctcc tcaactccag ggacaactcc catcccccca gtgctgacca 10321 ccaccgccac cacacctgca gccaccagca gcacagtgac tccctcctct gccctaggga 10381 ccacccacac acccccagtg ccgaacacca cggccaccac acacgggcgg tccctgcccc 10441 ccagcagtcc ccacacggtg cgcacagcct ggacttcggc cacctcgggc atcttgggca 10501 ccacccacat cacagagcct tccacggtga cttcccacac cccagcagca accaccagta 10561 ccacccagca ctcgactcca gccctgtcca gccctcaccc tagcagcagg accaccgagt 10621 cacccccttc tccagggacg accaccccgg gccacaccag gggcacctcc aggaccacag 10681 ccacagccac acccagcaag acccgcacct cgaccctgct gcccagcagc cccacatcgg 10741 cccccataac cacggtggtg accacgggct gtgagcccca gtgtgcctgg tcagagtggc 10801 tggactacag ctaccccatg ccggggccct ctggcgggga ctttgacacc tactccaaca 10861 tccgtgcggc cggaggggca gtctgtgagc agcccctggg cctcgagtgc cgtgcccagg 10921 cccagcctgg tgtccccctg cgggagttgg gccaggtcgt ggaatgcagc ctggactttg 10981 gcctggtctg caggaaccgt gagcaggtgg ggaagttcaa gatgtgcttc aactatgaaa 11041 tccgtgtgtt ctgctgcaac tacggccact gccccagcac cccggccacc agctctacgg 11101 ccacgccctc ctcaactccg gggacgacct ggatcctcac aaagctgacc acaacagcca 11161 ctacgactga gtccactgga tccacggcca ccccgtcctc caccccaggg accacctgga 11221 tcctcacaga gccgagcact acagccaccg tgacggtgcc caccggatcc acggccaccg 11281 cctcctccac ccaggcaact gctggcaccc cacatgtgag caccacggcc acgacaccca 11341 cagtcaccag ctccaaagcc actcccttct ccagtccagg gactgcaacc gcccttccag 11401 cactgagaag cacagccacc acacccacag ctaccagctt tacagccatc ccctcctcct 11461 ccctgggcac cacctggacc cgcctatcac agaccaccac acccacggcc accatgtcca 11521 cagccacacc ctcctccact ccagagactg cccacacctc cacagtgctt accaccacgg 11581 ccaccacaac cagggccacc ggctctgtgg ccaccccctc ttccacccca ggaacagctc 11641 acactaccaa agtgccgact accacaacca cgggcttcac agtcaccccc tcctccagcc 11701 cagggacggc acgcacgcct ccagtgtgga tcagcacaac caccacaccc acaaccagtg 11761 gctccacggt gaccccctcc tccgtcccgg ggaccaccca cacccccaca gtgctgacca 11821 ccaccaccac aactgtggcc actggttcta tggcaacacc ctcctctagc acacagacca 11881 gtggtactcc cccatcactg atcaccacgg ccactacgat cacggccacc ggctccacca 11941 ccaacccctc ctcaactcca gggacaacac ctatcccccc agtgctgacc accaccgcca 12001 ccacacctgc agccaccagc agcacagtga ctccctcctc tgccctaggg accacccaca 12061 cacccccagt gccgaacacc acggccacca cacacgggcg atccctgtcc cccagcagtc 12121 cccacacggt gcgcacagcc tggacttcgg ccacctcagg caccttgggc accacccaca 12181 tcacagagcc ttccacgggg acttcccaca ccccagcagc aaccaccggt accacccagc 12241 actcgactcc agccctgtcc agccctcacc ctagcagcag gaccaccgag tcaccccctt 12301 ccccagggac gaccaccccg ggccacacca cggccacctc caggaccacg gccacggcca 12361 cacccagcaa gacccgcacc tcgaccctgc tgcccagcag ccccacatcg gcccccataa 12421 ccacggtggt gaccacgggc tgtgagcccc agtgtgcctg gtcagagtgg ctggactaca 12481 gctaccccat gccggggccc tctggcgggg actttgacac ctactccaac atccgtgcgg 12541 ccggaggggc cgtctgtgag cagcccctgg gcctcgagtg ccgtgcccag gcccagcctg 12601 gtgtccccct gggggagttg ggccaggtcg tggaatgcag cctggacttt ggcctggtct 12661 gcaggaaccg tgagcaggtg gggaagttca agatgtgctt caactatgaa atccgtgtgt 12721 tctgctgcaa ctacggccac tgccccagca ccccggccac cagctctacg gccatgccct 12781 cctccactcc ggggacgacc tggatcctca cagagctgac cacaacagcc actacgactg 12841 catccactgg atccacggcc accccgtcct ccaccccggg aacagctccc cctcccaaag 12901 tgctgaccag cccggccacc acacccacag ccaccagttc caaagccact tcctcctcca 12961 gtccaaggac tgcaaccacc cttccagtgc tgacaagcac agccaccaaa tccacagcta 13021 ccagcgttac acccatcccc tcctccaccc ttgggaccac cgggaccctc ccagaacaga 13081 ccaccacacc cgtggccacc atgtccacaa tccacccctc ctccactccg gagaccaccc 13141 acacctccac agtgctgacc acgaaggcca ccacgacaag ggccaccagt tccacgtcca 13201 ccccctcctc cactccgggg acgacctgga tcctcacaga gctgaccaca gcagccacta 13261 caactgcagc cactggcccc acggccaccc cgtcctccac cccagggacc acctggatcc 13321 tcacagagct gaccacaaca gccactacga ctgcgtccac tggatccacg gccaccccgt 13381 cctccacccc agggaccacc tggatcctca cagagccgag cactacagcc accgtgacgg 13441 tgcccaccgg atccacggcc accgcctcct ccacccaggc aactgctggc accccacatg 13501 tgagcaccac ggccacgaca cccacagtca ccagctccaa agccactccc tcctccagtc 13561 cagggactgc aactgccctt ccagcactga gaagcacagc caccacaccc acagctacca 13621 gctttacagc catcccctcc tcctccctgg gcaccacctg gacccgccta tcacagacca 13681 ccacacccac ggccaccatg tccacagcca caccctcctc cactccagag actgtccaca 13741 cctccacagt gcttaccgcc acggccacca caaccggggc caccggctct gtggccaccc 13801 cctcctccac cccaggaaca gctcacacta ccaaagtgcc gactaccaca accacgggct 13861 tcacagccac cccctcctcc agcccaggga cggcactcac gcctccagtg tggatcagca 13921 caaccaccac acccacaacc accacaccca caaccagtgg ctccacggtg accccctcct 13981 ccatcccggg gaccacccac accgccagag tgctgaccac caccaccaca actgtggcca 14041 ctggttctat ggcaacaccc tcctctagca cacagaccag tggtactccc ccatcactga 14101 ccaccacggc cactacgatc acggccaccg gctccaccac caacccctcc tcaactccag 14161 ggacaacacc catcacccca gtgctgacca gcacggccac cacacccgca gccaccagct 14221 ccaaagccac ttcctcctcc agtccaagga ctgcaaccac ccttccagtg ctgacaagca 14281 cagccacaaa atccacagct accagcttta cacccatccc ctcctccacc ctgtggacca 14341 cgtggaccgt cccagcacag accaccacac ccatgtccac catgtccaca atccacacct 14401 cctctactcc agagaccacc cacacctcca cagtgctgac caccacagcc accatgacaa 14461 gggccaccaa ttccacggcc acaccctcct ccactctggg gacgacccgg atcctcactg 14521 agctgaccac aacagccact acaactgcag ccactggatc cacggccacc ctgtcctcca 14581 ccccagggac cacctggatc ctcacagagc cgagcactat agccaccgtg atggtgccca 14641 ccggttccac ggccaccgcc tcctccactc tgggaacagc tcacaccccc aaagtggtga 14701 ccaccatggc cactatgccc acagccactg cctccacggt tcccagctcg tccaccgtgg 14761 ggaccacccg cacccctgca gtgctcccca gcagcctgcc aaccttcagc gtgtccactg 14821 tgtcctcctc agtcctcacc accctgagac ccactggctt ccccagctcc cacttctcta 14881 ctccctgctt ctgcagggca tttggacagt ttttctcgcc cggggaagtc atctacaata 14941 agaccgaccg agccggctgc catttctacg cagtgtgcaa tcagcactgt gacattgacc 15001 gcttccaggg cgcctgtccc acctccccac cgccagtgtc ctccgccccg ctgtcctcgc 15061 cctcccctgc ccctggctgt gacaatgcca tccctctccg gcaggtgaat gagacctgga 15121 ccctggagaa ctgcacggtg gccaggtgcg tgggtgacaa ccgtgtcgtc ctgctggacc 15181 caaagcctgt ggccaacgtc acctgcgtga acaagcacct gcccatcaaa gtgtcggacc 15241 cgagccagcc ctgtgacttc cactatgagt gcgagtgcat ctgcagcatg tggggcggct 15301 cccactattc cacctttgac ggcacctctt acaccttccg gggcaactgc acctatgtcc 15361 tcatgagaga gatccatgca cgctttggga atctcagcct ctacctggac aaccactact 15421 gcacggcctc tgccactgcc gctgccgccc gctgcccccg cgccctcagc atccactaca 15481 agtccatgga tatcgtcctc actgtcacca tggtgcatgg gaaggaggag ggcctgatcc 15541 tgtttgacca aattccggtg agcagcggtt tcagcaagaa cggcgtgctt gtgtctgtgc 15601 tggggaccac caccatgcgt gtggacattc ctgccctggg cgtgagcgtc accttcaatg 15661 gccaagtctt ccaggcccgg ctgccctaca gcctcttcca caacaacacc gagggccagt 15721 gcggcacctg caccaacaac cagagggacg actgtctcca gcgggacgga accactgccg 15781 ccagttgcaa ggacatggcc aagacgtggc tggtccccga cagcagaaag gatggctgct 15841 gggccccgac tggcacaccc cccactgcca gccccgcagc cccggtgtct agcacaccca 15901 cccccacccc atgcccacca cagccgctct gtgatctgat gctgagccag gtctttgctg 15961 agtgccacaa ccttgtgccc ccgggcccat tcttcaacgc ctgcatcagc gaccactgca 16021 ggggccgcct tgaggtgccc tgccagagcc tggaggctta cgcagagctc tgccgcgccc 16081 ggggagtgtg cagtgactgg cgaggtgcaa ccggtggcct gtgcgacctc acctgcccac 16141 ccaccaaagt gtacaagcca tgcggcccca tacagcctgc cacctgcaac tctaggaacc 16201 agagcccaca gctggagggg atggcggagg gctgcttctg ccctgaggac cagatcctct 16261 tcaacgcaca catgggcatc tgcgtgcagg cctgcccctg cgtgggaccc gatgggtttc 16321 ctaaatttcc cggggagcgg tgggtcagca actgccagtc ctgcgtgtgt gacgagggtt 16381 cagtgtcggt gcagtgcaag cccctgccct gtgacgccca gggtcagccc ccgccgtgca 16441 accgtcccgg cttcgtaacc gtgaccaggc cccgggccga gaacccctgc tgccccgaga 16501 cggtgtgcgt gtgcaacaca accacctgcc cccagagcct gcctgtgtgc ccgccagggc 16561 aggagtccat ctgcacccag gaggagggcg actgctgtcc caccttccgc tgcagacctc 16621 agctgtgttc gtacaatggc accttctacg gggttggtgc aaccttccca ggcgcccttc 16681 cctgccacat gtgtacctgc ctctctgggg acacccagga cccaacggtg caatgtcagg 16741 aggatgcctg caacaatact acctgtcccc agggctttga gtacaagaga gtggccgggc 16801 agtgctgtgg ggagtgcgtc cagaccgcct gcctcacgcc cgatggccag ccagtccagc 16861 tgaatgaaac ctgggtcaac agccatgtgg acaactgcac cgtgtacctc tgtgaggctg 16921 agggtggagt ccatttgctg accccacagc ctgcatcctg cccagatgtg tccagctgca 16981 gggggagcct caggaaaacc ggctgctgct actcctgtga ggaggactcc tgtcaagtcc 17041 gcatcaacac gaccatcctg tggcaccagg gctgcgagac cgaggtcaac atcaccttct 17101 gcgagggctc ctgccccgga gcgtccaagt actcagcaga ggcccaggcc atgcagcacc 17161 agtgcacctg ctgccaggag aggcgggtcc acgaggagac ggtgcccttg cactgtccta 17221 acggctcagc catcctgcac acctacaccc acgtggatga gtgtggctgc acgcccttct 17281 gtgtccctgc gcccatggct cccccacaca cccgtggctt cccggcccag gaggccactg 17341 ctgtctgaga acgttctgcc tccatcccca tgctctgtcc acctggagcc aggatgtgca 17401 ttgtctgatc atgaaaacct tgggcctcct ctgcggagcc ccccggcctg tgtgtggcac 17461 cccgcgctcc gtgctcctgc tgcccacccc gtgggtgaaa ccggccccag aagggtgagg 17521 ggccagcagg acccctttcg ggagggcgcc actcaggagt cctaccctgg gagagcctgt 17581 ggcccacctt ggccttgccc ctccctgatg tcactgggac gccctggaac aaactaagca 17641 tgtgcgggcc tatgtgtccc tgccacggcc ggagcgcccg cgcagcacgg attccagctg 17701 gccacgtccg gccgctgggg cagacaggct ggtccaggca aggccagctg ctgccaggaa 17761 gctgcgacag gcaaggcggc cgcctgtcca tgcctgctgc agggtaactc agggctgagg 17821 tcgcaacggc caggtcagag aggggtcagc atcccaaagc cccctctgct caacccagcc 17881 cagttttgca aataaaccct gagcattgag tacgtt 

In some embodiments of the methods of the disclosure, the wild type human MUC5B gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_002449.2):

(SEQ ID NO: 12)     1 mgapsacrtl vlalaamlvv pqaetqgpve pswenaghtm dggaptsspt rrvsfvppvt   61 vfpslspinp ahngrvcstw gdfhyktfdg dvfrfpglcn yvfsehcraa yedfnvqlrr  121 glvgsrpvvt rvvikaqglv leasngsvli ngqreelpys rtgllveqsg dyikvsirlv  181 ltflwngeds alleldpkya nqtcglcgdf nglpafnefy ahnarltplq fgnlqkldgp  241 teqcpdplpl pagnctdeeg ichrtllgpa faechalvds taylaacaqd lcrcptcpca  301 tfveysrqca haggqprnwr cpelcprtcp lnmqhqecgs pctdtcsnpq raqlcedhcv  361 dgcfcppgtv lddithsgcl plgqcpcthg grtyspgtsf nttcssctcs gglwqcqdlp  421 cpgtcsvqgg ahistydekl ydlhgdcsyv lskkcadssf tvlaelrkcg ltdnenclka  481 vtlsldggdt airvqadggv flnsiytqlp lsaanitlft pssffivvqt glglqllvql  541 vplmqvfvrl dpahqgqmcg lcgnfnqnqa ddftalsgvv eatgaafant wkaqaacana  601 rnsfedpcsl svenenyarh wcsrltdpns afsrchsiin pkpfhsncmf dtcncersed  661 cicaalssyv hacaakgvql sdwrdgvctk ymqncpksqr yayvvdacqp tcrglseadv  721 tcsysfvpvd gctcpagtfl ndagacvpaq ecpcyahgtv lapgevvhde gavcsctggk  781 lsclgaslqk stgcaapmvy ldcsnssagt pgaeclrsch tldvgcfsth cvsgcvcppg  841 lvsdgsggci aeedcpcvhn eatykpgeti rvdcntctcr nrrwecshrl clgtcvaygd  901 ghfitfdgdr ysfegsceyi laqdycgdnt thgtfrivte nipcgttgtt cskaiklfve  961 syelilqegt fkavargpgg dppykirymg iflviethgm ayswdrktsv firlhqdykg 1021 rvcglcgnfd dnaindfatr srsvvgdale fgnswklsps cpdalapkdp ctanpfrksw 1081 aqkqcsilhg ptfaacrsqv dstkyyeacv ndacacdsgg dcecfctava ayaqachdag 1141 lcvswrtpdt cplfcdfynp hggcewhyqp cgapclktcr npsghclvdl pglegcypkc 1201 ppsqpffned qmkcvaqcgc ydkdgnyydv garvptaenc qscnctpsgi qcahsleact 1261 ctyedrtysy qdviynttdg lgacliaicg sngtiirkav acpgtpattp ftfttawvph 1321 sttspalpvs tvcvrevcrw sswynghrpe pglgggdfet fenlrqrgyq vcpvladiec 1381 raaqlpdmpl eelgqqvdcd rmrglmcans qqspplchdy elrvlcceyv pcgpspapgt 1441 spqpslsast epavptptqt tatekttlwv tpsirstaal tsqtgsssgp vtvtpsapgt 1501 ttcqprcqwt ewfdedypks eqlggdvesy dkiraagghl cqqpkdiecq aesfpnwtla 1561 qvgqkvhcdv hfglvcrnwe qegvfkmcyn yrirvlccsd dhcrgrattp pptteletat 1621 ttttqalfst pqptsspglt rappasttav ptlsegltsp rytstlgtat tggpttpags 1681 teptvpgvat stlptrsalp gttgslgtwr psqpptlapt tmatsrarpt gtastaskep 1741 lttslaptlt selstsqaet stprtettms pltntttsqg ttrcqpkcew tewfdvdfpt 1801 sgvaggdmet feniraaggk mcwapksiec raenypevsi dqvgqvltcs letgltckne 1861 dqtgrfnmcf nynvrvlccd dyshcpstpa tsstatpsst pgttwiltkp tttatttast 1921 gstatptstl rtapppkvlt ttattptvts skatpssspg tatalpalrs tattptatsv 1981 tpipssslgt twtrlsqttt ptatmstatp sstpetahts tvltatattt gatgsvatps 2041 stpgtahttk vptttttgft atpssspgta ltppvwistt ttpttrgstv tpssipgtth 2101 tatvlttttt tvatgsmatp ssstqtsgtp psltttatti tatgsttnps stpgttpipp 2161 vltttattpa atsntvtpss algtthtppv pntmatthgr slppssphtv rtawtsatsg 2221 ilgtthitep stvtshtlaa ttgttqhstp alssphpssr ttesppspgt ttpghttats 2281 rttatatpsk trtstllpss ptsapittvv tmgcepqcaw sewldysypm pgpsggdfdt 2341 ysniraagga vceqplglec raqaqpgvpl relgqvvecs ldfglvcrnr eqvgkfkmcf 2401 nyeirvfccn yghcpstpat sstampsstp gttwiltelt ttatttestg statpsstpg 2461 ttwiltepst tatvtvptgs tatasstqat agtphvstta ttptvtsska tpfsspgtat 2521 alpalrstat tptatsftai pssslgttwt rlsqtttpta tmstatpsst petvhtstvl 2581 tttatttgat gsvatpsstp gtahttkvlt ttttgftatp ssspgtartl pvwisttttp 2641 ttrgstvtps sipgtthtpt vlttttttva tgsmatpsss tqtsgtppsl tttattitat 2701 gsttnpsstp gttpippvlt ttattpaats stvtpssalg tthtppvpnt tatthgrsls 2761 pssphtvrta wtsatsgtlg tthitepstg tshtpaattg ttqhstpals sphpssrtte 2821 sppspgtttp ghtratsrtt atatpsktrt stllpsspts apittvvtmg cepqcawsew 2881 ldysypmpgp sggdfdtysn iraaggavce qplglecraq aqpgvplrel gqvvecsldf 2941 glvcrnreqv gkfkmcfnye irvfccnygh cpstpatsst atpsstpgtt wilteqttaa 3001 tttattgsta ipsstpgtap ppkvltstat tptatsskat ssssprtatt 1pvltstatk 3061 statsftpip sftlgttgtl peqtttpmat mstihpsstp etthtstvlt tkatttrats 3121 smstpsstpg ttwilteltt aatttaatgp tatpsstpgt twiltepstt atvtvptgst 3181 atasstrata gtlkvltsta ttptvissra tpssspgtat alpalrstat tptatsvtai 3241 pssslgtawt rlsqtttpta tmstatpsst petvhtstvl tttttttrat gsvatpsstp 3301 gtahttkvpt ttttgftatp ssspgtaltp pvwisttttp ttrgstvtps sipgtthtat 3361 vlttttttva tgsmatpsss tqtsgtppsl tttattitat gsttnpsstp gttpippvlt 3421 ttattpaats stvtpssalg tthtppvpnt tatthgrslp pssphtvrta wtsatsgilg 3481 tthitepstv tshtpaatts ttqhstpals sphpssrtte sppspgtttp ghtrgtsrtt 3541 atatpsktrt stllpsspts apittvvttg cepqcawsew ldysypmpgp sggdfdtysn 3601 iraaggavce qplglecraq aqpgvplrel gqvvecsldf glvcrnreqv gkfkmcfnye 3661 irvfccnygh cpstpatsst atpsstpgtt wiltklttta tttestgsta tpsstpgttw 3721 iltepsttat vtvptgstat asstqatagt phvsttattp tvtsskatpf sspgtatalp 3781 alrstattpt atsftaipss slgttwtrls qtttptatms tatpsstpet ahtstvlttt 3841 atttratgsv atpsstpgta httkvptttt tgftvtpsss pgtartppvw isttttptts 3901 gstvtpssvp gtthtptvlt tttttvatgs matpssstqt sgtppslitt attitatgst 3961 tnpsstpgtt pippvlttta ttpaatsstv tpssalgtth tppvpnttat thgrslspss 4021 phtvrtawts atsgtlgtth itepstgtsh tpaattgttq hstpalssph pssrttespp 4081 spgtttpght tatsrttata tpsktrtstl 1pssptsapi ttvvttgcep qcawsewldy 4141 sypmpgpsgg dfdtysnira aggavceqpl glecraqaqp gvplgelgqv vecsldfglv 4201 crnreqvgkf kmcfnyeiry fccnyghcps tpatsstamp sstpgttwil teltttattt 4261 astgstatps stpgtapppk vltspattpt atsskatsss sprtattlpv ltstatksta 4321 tsvtpipsst lgttgtlpeq tttpvatmst ihpsstpett htstvlttka tttratssts 4381 tpsstpgttw iltelttaat ttaatgptat psstpgttwi lteltttatt tastgstatp 4441 sstpgttwil tepsttatvt vptgstatas stqatagtph vsttattptv tsskatpsss 4501 pgtatalpal rstattptat sftaipsssl gttwtrlsqt ttptatmsta tpsstpetvh 4561 tstvltatat ttgatgsvat psstpgtaht tkvptttttg ftatpssspg taltppvwis 4621 ttttpttttp ttsgstvtps sipgtthtar vlttttttva tgsmatpsss tqtsgtppsl 4681 tttattitat gsttnpsstp gttpitpvlt stattpaats skatsssspr tattlpvlts 4741 tatkstatsf tpipsstlwt twtvpaqttt pmstmstiht sstpetthts tvltttatmt 4801 ratnstatps stlgttrilt eltttattta atgstatlss tpgttwilte pstiatvmvp 4861 tgstatasst lgtahtpkvv ttmatmptat astvpssstv gttrtpavlp sslptfsyst 4921 vsssvlttlr ptgfpsshfs tpcfcrafgq ffspgeviyn ktdragchfy avcnqhcdid 4981 rfqgacptsp ppvssaplss pspapgcdna iplrqvnetw tlenctvarc vgdnrvvlld 5041 pkpvanvtcv nkhlpikvsd psqpcdfhye cecicsmwgg shystfdgts ytfrgnctyv 5101 lmreiharfg nlslyldnhy ctasataaaa rcpralsihy ksmdivltvt mvhgkeegli 5161 lfdqipvssg fskngvlvsv lgtttmrvdi palgvsvtfn gqvfqarlpy slfhnntegq 5221 cgtctnnqrd dclqrdgtta asckdmaktw lvpdsrkdgc waptgtppta spaapvsstp 5281 tptpcppqpl cdlmlsqvfa echnlvppgp ffnacisdhc rgrlevpcqs leayaelcra 5341 rgvcsdwrga tgglcdltcp ptkvykpcgp iqpatcnsrn qspqlegmae gcfcpedqil 5401 fnahmgicvq acpcvgpdgf pkfpgerwvs ncqscvcdeg sysvqckplp cdaqgqpppc 5461 nrpgfvtvtr praenpccpe tvcvcntttc pqslpvcppg qesictqeeg dccptfrcrp 5521 qlcsyngtfy gvgatfpgal pchmctclsg dtqdptvqcq edacnnttcp qgfeykrvag 5581 qccgecvqta cltpdgqpvq lnetwvnshv dnctvylcea eggvhlltpq pascpdvssc 5641 rgslrktgcc ysceedscqv rinttilwhq gcetevnitf cegscpgask ysaeaqamqh 5701 qctccqerry heetvplhcp ngsailhtyt hvdecgctpf cvpapmapph trgfpaqeat 5761 av 

In some embodiments of the methods of the disclosure, the wild type human TERT gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_198253.2, transcript variant 1):

(SEQ ID NO: 13)    1 caggcagcgc tgcgtcctgc tgcgcacgtg ggaagccctg gccccggcca cccccgcgat   61 gccgcgcgct ccccgctgcc gagccgtgcg ctccctgctg cgcagccact accgcgaggt  121 gctgccgctg gccacgttcg tgcggcgcct ggggccccag ggctggcggc tggtgcagcg  181 cggggacccg gcggctttcc gcgcgctggt ggcccagtgc ctggtgtgcg tgccctggga  241 cgcacggccg ccccccgccg ccccctcctt ccgccaggtg tcctgcctga aggagctggt  301 ggcccgagtg ctgcagaggc tgtgcgagcg cggcgcgaag aacgtgctgg ccttcggctt  361 cgcgctgctg gacggggccc gcgggggccc ccccgaggcc ttcaccacca gcgtgcgcag  421 ctacctgccc aacacggtga ccgacgcact gcgggggagc ggggcgtggg ggctgctgct  481 gcgccgcgtg ggcgacgacg tgctggttca cctgctggca cgctgcgcgc tctttgtgct  541 ggtggctccc agctgcgcct accaggtgtg cgggccgccg ctgtaccagc tcggcgctgc  601 cactcaggcc cggcccccgc cacacgctag tggaccccga aggcgtctgg gatgcgaacg  661 ggcctggaac catagcgtca gggaggccgg ggtccccctg ggcctgccag ccccgggtgc  721 gaggaggcgc gggggcagtg ccagccgaag tctgccgttg cccaagaggc ccaggcgtgg  781 cgctgcccct gagccggagc ggacgcccgt tgggcagggg tcctgggccc acccgggcag  841 gacgcgtgga ccgagtgacc gtggtttctg tgtggtgtca cctgccagac ccgccgaaga  901 agccacctct ttggagggtg cgctctctgg cacgcgccac tcccacccat ccgtgggccg  961 ccagcaccac gcgggccccc catccacatc gcggccacca cgtccctggg acacgccttg 1021 tcccccggtg tacgccgaga ccaagcactt cctctactcc tcaggcgaca aggagcagct 1081 gcggccctcc ttcctactca gctctctgag gcccagcctg actggcgctc ggaggctcgt 1141 ggagaccatc tttctgggtt ccaggccctg gatgccaggg actccccgca ggttgccccg 1201 cctgccccag cgctactggc aaatgcggcc cctgtttctg gagctgcttg ggaaccacgc 1261 gcagtgcccc tacggggtgc tcctcaagac gcactgcccg ctgcgagctg cggtcacccc 1321 agcagccggt gtctgtgccc gggagaagcc ccagggctct gtggcggccc ccgaggagga 1381 ggacacagac ccccgtcgcc tggtgcagct gctccgccag cacagcagcc cctggcaggt 1441 gtacggcttc gtgcgggcct gcctgcgccg gctggtgccc ccaggcctct ggggctccag 1501 gcacaacgaa cgccgcttcc tcaggaacac caagaagttc atctccctgg ggaagcatgc 1561 caagctctcg ctgcaggagc tgacgtggaa gatgagcgtg cgggactgcg cttggctgcg 1621 caggagccca ggggttggct gtgttccggc cgcagagcac cgtctgcgtg aggagatcct 1681 ggccaagttc ctgcactggc tgatgagtgt gtacgtcgtc gagctgctca ggtctttctt 1741 ttatgtcacg gagaccacgt ttcaaaagaa caggctcttt ttctaccgga agagtgtctg 1801 gagcaagttg caaagcattg gaatcagaca gcacttgaag agggtgcagc tgcgggagct 1861 gtcggaagca gaggtcaggc agcatcggga agccaggccc gccctgctga cgtccagact 1921 ccgcttcatc cccaagcctg acgggctgcg gccgattgtg aacatggact acgtcgtggg 1981 agccagaacg ttccgcagag aaaagagggc cgagcgtctc acctcgaggg tgaaggcact 2041 gttcagcgtg ctcaactacg agcgggcgcg gcgccccggc ctcctgggcg cctctgtgct 2101 gggcctggac gatatccaca gggcctggcg caccttcgtg ctgcgtgtgc gggcccagga 2161 cccgccgcct gagctgtact ttgtcaaggt ggatgtgacg ggcgcgtacg acaccatccc 2221 ccaggacagg ctcacggagg tcatcgccag catcatcaaa ccccagaaca cgtactgcgt 2281 gcgtcggtat gccgtggtcc agaaggccgc ccatgggcac gtccgcaagg ccttcaagag 2341 ccacgtctct accttgacag acctccagcc gtacatgcga cagttcgtgg ctcacctgca 2401 ggagaccagc ccgctgaggg atgccgtcgt catcgagcag agctcctccc tgaatgaggc 2461 cagcagtggc ctcttcgacg tcttcctacg cttcatgtgc caccacgccg tgcgcatcag 2521 gggcaagtcc tacgtccagt gccaggggat cccgcagggc tccatcctct ccacgctgct 2581 ctgcagcctg tgctacggcg acatggagaa caagctgttt gcggggattc ggcgggacgg 2641 gctgctcctg cgtttggtgg atgatttctt gttggtgaca cctcacctca cccacgcgaa 2701 aaccttcctc aggaccctgg tccgaggtgt ccctgagtat ggctgcgtgg tgaacttgcg 2761 gaagacagtg gtgaacttcc ctgtagaaga cgaggccctg ggtggcacgg cttttgttca 2821 gatgccggcc cacggcctat tcccctggtg cggcctgctg ctggataccc ggaccctgga 2881 ggtgcagagc gactactcca gctatgcccg gacctccatc agagccagtc tcaccttcaa 2941 ccgcggcttc aaggctggga ggaacatgcg tcgcaaactc tttggggtct tgcggctgaa 3001 gtgtcacagc ctgtttctgg atttgcaggt gaacagcctc cagacggtgt gcaccaacat 3061 ctacaagatc ctcctgctgc aggcgtacag gtttcacgca tgtgtgctgc agctcccatt 3121 tcatcagcaa gtttggaaga accccacatt tttcctgcgc gtcatctctg acacggcctc 3181 cctctgctac tccatcctga aagccaagaa cgcagggatg tcgctggggg ccaagggcgc 3241 cgccggccct ctgccctccg aggccgtgca gtggctgtgc caccaagcat tcctgctcaa 3301 gctgactcga caccgtgtca cctacgtgcc actcctgggg tcactcagga cagcccagac 3361 gcagctgagt cggaagctcc cggggacgac gctgactgcc ctggaggccg cagccaaccc 3421 ggcactgccc tcagacttca agaccatcct ggactgatgg ccacccgccc acagccaggc 3481 cgagagcaga caccagcagc cctgtcacgc cgggctctac gtcccaggga gggaggggcg 3541 gcccacaccc aggcccgcac cgctgggagt ctgaggcctg agtgagtgtt tggccgaggc 3601 ctgcatgtcc ggctgaaggc tgagtgtccg gctgaggcct gagcgagtgt ccagccaagg 3661 gctgagtgtc cagcacacct gccgtcttca cttccccaca ggctggcgct cggctccacc 3721 ccagggccag cttttcctca ccaggagccc ggcttccact ccccacatag gaatagtcca 3781 tccccagatt cgccattgtt cacccctcgc cctgccctcc tttgccttcc acccccacca 3841 tccaggtgga gaccctgaga aggaccctgg gagctctggg aatttggagt gaccaaaggt 3901 gtgccctgta cacaggcgag gaccctgcac ctggatgggg gtccctgtgg gtcaaattgg 3961 ggggaggtgc tgtgggagta aaatactgaa tatatgagtt tttcagtttt gaaaaaaa 

In some embodiments of the methods of the disclosure, the wild type human TERT gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_937983.2, transcript variant 1):

(SEQ ID NO: 14)    1 mpraprcrav rsllrshyre vlplatfvrr lgpqgwrlvq rgdpaafral vaqclvcvpw   61 darpppaaps frqvsclkel varvlqrlce rgaknvlafg falldgargg ppeafttsvr  121 sylpntvtda lrgsgawgll lrrvgddvlv hllarcalfv lvapscayqv cgpplyqlga  181 atqarpppha sgprrrlgce rawnhsvrea gvplglpapg arrrggsasr slplpkrprr  241 gaapepertp vgqgswahpg rtrgpsdrgf cvvsparpae eatslegals gtrhshpsvg  301 rqhhagppst srpprpwdtp cppvyaetkh flyssgdkeq lrpsfllssl rpsltgarrl  361 vetiflgsrp wmpgtprrlp rlpqrywqmr plflellgnh aqcpygvllk thcplraavt  421 paagvcarek pqgsvaapee edtdprrlvq llrqhsspwq vygfvraclr rlvppglwgs  481 rhnerrflrn tkkfislgkh aklslqeltw kmsvrdcawl rrspgvgcvp aaehrlreei  541 lakflhwlms vyvvellrsf fyvtettfqk nrlffyrksv wsklqsigir qhlkrvqlre  601 lseaevrqhr earpalltsr lrfipkpdgl rpivnmdyvv gartfrrekr aerltsrvka  661 lfsvinyera rrpgllgasv lglddihraw rtfvlrvraq dpppelyfvk vdvtgaydti  721 pqdrltevia siikpqntyc vrryavvqka ahghvrkafk shvstltdlq pymrqfvahl  781 qetsplrdav vieqssslne assglfdvfl rfmchhavri rgksyvqcqg ipqgsilstl  841 lcslcygdme nklfagirrd glllrlvddf llvtphltha ktflrtivrg vpeygcvvnl  901 rktvvnfpve dealggtafv qmpahglfpw cglildtrtl evqsdyssya rtsirasltf  961 nrgfkagrnm rrklfgvlrl kchslfldlq vnslqtvctn iykilllqay rfhacvlqlp 1021 fhqqvwknpt fflrvisdta slcysilkak nagmslgakg aagplpseav qwlchqafll 1081 kltrhrvtyv pllgslrtaq tqlsrklpgt tltaleaaan palpsdfkti ld 

In some embodiments of the methods of the disclosure, the wild type human TERT gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_001193376.1, transcript variant 2):

(SEQ ID NO: 15)    1 caggcagcgc tgcgtcctgc tgcgcacgtg ggaagccctg gccccggcca cccccgcgat   61 gccgcgcgct ccccgctgcc gagccgtgcg ctccctgctg cgcagccact accgcgaggt  121 gctgccgctg gccacgttcg tgcggcgcct ggggccccag ggctggcggc tggtgcagcg  181 cggggacccg gcggctttcc gcgcgctggt ggcccagtgc ctggtgtgcg tgccctggga  241 cgcacggccg ccccccgccg ccccctcctt ccgccaggtg tcctgcctga aggagctggt  301 ggcccgagtg ctgcagaggc tgtgcgagcg cggcgcgaag aacgtgctgg ccttcggctt  361 cgcgctgctg gacggggccc gcgggggccc ccccgaggcc ttcaccacca gcgtgcgcag  421 ctacctgccc aacacggtga ccgacgcact gcgggggagc ggggcgtggg ggctgctgct  481 gcgccgcgtg ggcgacgacg tgctggttca cctgctggca cgctgcgcgc tctttgtgct  541 ggtggctccc agctgcgcct accaggtgtg cgggccgccg ctgtaccagc tcggcgctgc  601 cactcaggcc cggcccccgc cacacgctag tggaccccga aggcgtctgg gatgcgaacg  661 ggcctggaac catagcgtca gggaggccgg ggtccccctg ggcctgccag ccccgggtgc  721 gaggaggcgc gggggcagtg ccagccgaag tctgccgttg cccaagaggc ccaggcgtgg  781 cgctgcccct gagccggagc ggacgcccgt tgggcagggg tcctgggccc acccgggcag  841 gacgcgtgga ccgagtgacc gtggtttctg tgtggtgtca cctgccagac ccgccgaaga  901 agccacctct ttggagggtg cgctctctgg cacgcgccac tcccacccat ccgtgggccg  961 ccagcaccac gcgggccccc catccacatc gcggccacca cgtccctggg acacgccttg 1021 tcccccggtg tacgccgaga ccaagcactt cctctactcc tcaggcgaca aggagcagct 1081 gcggccctcc ttcctactca gctctctgag gcccagcctg actggcgctc ggaggctcgt 1141 ggagaccatc tttctgggtt ccaggccctg gatgccaggg actccccgca ggttgccccg 1201 cctgccccag cgctactggc aaatgcggcc cctgtttctg gagctgcttg ggaaccacgc 1261 gcagtgcccc tacggggtgc tcctcaagac gcactgcccg ctgcgagctg cggtcacccc 1321 agcagccggt gtctgtgccc gggagaagcc ccagggctct gtggcggccc ccgaggagga 1381 ggacacagac ccccgtcgcc tggtgcagct gctccgccag cacagcagcc cctggcaggt 1441 gtacggcttc gtgcgggcct gcctgcgccg gctggtgccc ccaggcctct ggggctccag 1501 gcacaacgaa cgccgcttcc tcaggaacac caagaagttc atctccctgg ggaagcatgc 1561 caagctctcg ctgcaggagc tgacgtggaa gatgagcgtg cgggactgcg cttggctgcg 1621 caggagccca ggggttggct gtgttccggc cgcagagcac cgtctgcgtg aggagatcct 1681 ggccaagttc ctgcactggc tgatgagtgt gtacgtcgtc gagctgctca ggtctttctt 1741 ttatgtcacg gagaccacgt ttcaaaagaa caggctcttt ttctaccgga agagtgtctg 1801 gagcaagttg caaagcattg gaatcagaca gcacttgaag agggtgcagc tgcgggagct 1861 gtcggaagca gaggtcaggc agcatcggga agccaggccc gccctgctga cgtccagact 1921 ccgcttcatc cccaagcctg acgggctgcg gccgattgtg aacatggact acgtcgtggg 1981 agccagaacg ttccgcagag aaaagagggc cgagcgtctc acctcgaggg tgaaggcact 2041 gttcagcgtg ctcaactacg agcgggcgcg gcgccccggc ctcctgggcg cctctgtgct 2101 gggcctggac gatatccaca gggcctggcg caccttcgtg ctgcgtgtgc gggcccagga 2161 cccgccgcct gagctgtact ttgtcaaggt ggatgtgacg ggcgcgtacg acaccatccc 2221 ccaggacagg ctcacggagg tcatcgccag catcatcaaa ccccagaaca cgtactgcgt 2281 gcgtcggtat gccgtggtcc agaaggccgc ccatgggcac gtccgcaagg ccttcaagag 2341 ccacgtctct accttgacag acctccagcc gtacatgcga cagttcgtgg ctcacctgca 2401 ggagaccagc ccgctgaggg atgccgtcgt catcgagcag agctcctccc tgaatgaggc 2461 cagcagtggc ctcttcgacg tcttcctacg cttcatgtgc caccacgccg tgcgcatcag 2521 gggcaagtcc tacgtccagt gccaggggat cccgcagggc tccatcctct ccacgctgct 2581 ctgcagcctg tgctacggcg acatggagaa caagctgttt gcggggattc ggcgggacgg 2641 gctgctcctg cgtttggtgg atgatttctt gttggtgaca cctcacctca cccacgcgaa 2701 aaccttcctc agctatgccc ggacctccat cagagccagt ctcaccttca accgcggctt 2761 caaggctggg aggaacatgc gtcgcaaact ctttggggtc ttgcggctga agtgtcacag 2821 cctgtttctg gatttgcagg tgaacagcct ccagacggtg tgcaccaaca tctacaagat 2881 cctcctgctg caggcgtaca ggtttcacgc atgtgtgctg cagctcccat ttcatcagca 2941 agtttggaag aaccccacat ttttcctgcg cgtcatctct gacacggcct ccctctgcta 3001 ctccatcctg aaagccaaga acgcagggat gtcgctgggg gccaagggcg ccgccggccc 3061 tctgccctcc gaggccgtgc agtggctgtg ccaccaagca ttcctgctca agctgactcg 3121 acaccgtgtc acctacgtgc cactcctggg gtcactcagg acagcccaga cgcagctgag 3181 tcggaagctc ccggggacga cgctgactgc cctggaggcc gcagccaacc cggcactgcc 3241 ctcagacttc aagaccatcc tggactgatg gccacccgcc cacagccagg ccgagagcag 3301 acaccagcag ccctgtcacg ccgggctcta cgtcccaggg agggaggggc ggcccacacc 3361 caggcccgca ccgctgggag tctgaggcct gagtgagtgt ttggccgagg cctgcatgtc 3421 cggctgaagg ctgagtgtcc ggctgaggcc tgagcgagtg tccagccaag ggctgagtgt 3481 ccagcacacc tgccgtcttc acttccccac aggctggcgc tcggctccac cccagggcca 3541 gcttttcctc accaggagcc cggcttccac tccccacata ggaatagtcc atccccagat 3601 tcgccattgt tcacccctcg ccctgccctc ctttgccttc cacccccacc atccaggtgg 3661 agaccctgag aaggaccctg ggagctctgg gaatttggag tgaccaaagg tgtgccctgt 3721 acacaggcga ggaccctgca cctggatggg ggtccctgtg ggtcaaattg gggggaggtg 3781 ctgtgggagt aaaatactga atatatgagt ttttcagttt tgaaaaaaa

In some embodiments of the methods of the disclosure, the wild type human TERT gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_001180305.1, transcript variant 21:

(SEQ ID NO: 16)    1 mpraprcrav rsllrshyre vlplatfvrr lgpqgwrlvq rgdpaafral vaqclvcvpw   61 darpppaaps frqvsclkel varvlqrlce rgaknvlafg falldgargg ppeafttsvr  121 sylpntvtda lrgsgawgll lrrvgddvlv hllarcalfv lvapscayqv cgpplyqlga  181 atqarpppha sgprrrlgce rawnhsvrea gvplglpapg arrrggsasr slplpkrprr  241 gaapepertp vgqgswahpg rtrgpsdrgf cvvsparpae eatslegals gtrhshpsvg  301 rqhhagppst srpprpwdtp cppvyaetkh flyssgdkeq lrpsfllssl rpsltgarrl  361 vetiflgsrp wmpgtprrlp rlpqrywqmr plflellgnh aqcpygvllk thcplraavt  421 paagvcarek pqgsvaapee edtdprrlvq llrqhsspwq vygfvraclr rlvppglwgs  481 rhnerrflrn tkkfislgkh aklslqeltw kmsvrdcawl rrspgvgcvp aaehrlreei  541 lakflhwlms vyvvellrsf fyvtettfqk nrlffyrksv wsklqsigir qhlkrvqlre  601 lseaevrqhr earpalltsr lrfipkpdgl rpivnmdyvv gartfrrekr aerltsrvka  661 lfsvinyera rrpgllgasv lglddihraw rtfvlrvraq dpppelyfvk vdvtgaydti  721 pqdrltevia siikpqntyc vrryavvqka ahghvrkafk shvstltdlq pymrqfvahl  781 qetsplrdav vieqssslne assglfdvfl rfmchhavri rgksyvqcqg ipqgsilstl  841 lcslcygdme nklfagirrd glllrlvddf llvtphltha ktflsyarts irasltfnrg  901 fkagrnmrrk lfgvlrlkch slfldlqvns lqtvctniyk illlqayrfh acvlqlpfhq  961 qvwknptffl rvisdtaslc ysilkaknag mslgakgaag plpseavqwl chqafllklt 1021 rhrvtyvpll gslrtaqtql srklpgttlt aleaaanpal psdfktild 

In some embodiments of the methods of the disclosure, the wild type human FAM13A gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_014883.3, transcript variant 1):

(SEQ ID NO: 36) 1 atcaaatttc aactccaggc agtccttcca gccatgtggg ttcagcggaa agagaagcaa 61 aaccactctt cctaaaatgt tagaagctgc tcttcgctta ccttggggcc tttgcattgg 121 gagctgtttt tcacatcaaa gaatatgtgc tgaatggaat tttagtattt tgctgtcgtt 181 ttaatatttt cgtctggtct tcctcagttc ttccagacgc tttctgagag aatgggggca 241 ggagctctag ccatctgtca aagtaaagca gcggttcggc tgaaagaaga catgaaaaag 301 atagtggcag tgccattaaa tgaacagaag gattttacct atcagaagtt atttggagtc 361 agtctccaag aacttgaacg gcaggggctc accgagaatg gcattccagc agtagtgtgg 421 aatatagtgg aatatttgac gcagcatgga cttacccaag aaggtctttt tagggtgaat 481 ggtaacgtga aggtggtgga acaacttcga ctgaagttcg agagtggagt gcccgtggag 541 ctcgggaagg acggtgatgt ctgctcagca gccagtctgt tgaagctgtt tctgagggag 601 ctgcctgaca gtctgatcac ctcagcgttg cagcctcgat tcattcaact ctttcaggat 661 ggcagaaatg atgttcagga gagtagctta agagacttaa taaaagagct gccagacacc 721 cactactgcc tcctcaagta cctttgccag ttcttgacaa aagtagccaa gcatcatgtg 781 cagaatcgca tgaatgttca caatctcgcc actgtatttg ggccaaattg ctttcatgtg 841 ccacctgggc ttgaaggcat gaaggaacag gacctgtgca acaagataat ggctaaaatt 901 ctagaaaatt acaataccct gtttgaagta gagtatacag aaaatgatca tctgagatgt 961 gaaaacctgg ctaggcttat catagtaaaa gaggtctatt ataagaactc cctgcccatc 1021 cttttaacaa gaggcttaga aagagacatg ccaaaaccac ctccaaaaac caagatccca 1081 aaatccagga gtgagggatc tattcaggcc cacagagtac tgcaaccaga gctatctgat 1141 ggcattcctc agctcagctt gcggctaagt tatagaaaag cctgcttgga agacatgaat 1201 tcagcagagg gtgctattag tgccaagttg gtacccagtt cacaggaaga tgaaagacct 1261 ctgtcacctt tctatttgag tgctcatgta ccccaagtca gcaatgtgtc tgcaaccgga 1321 gaactcttag aaagaaccat ccgatcagct gtagaacaac atctttttga tgttaataac 1381 tctggaggtc aaagttcaga ggactcagaa tctggaacac tatcagcatc ttctgccaca 1441 tctgccagac agcgccgccg ccagtccaag gagcaggatg aagttcgaca tgggagagac 1501 aagggactta tcaacaaaga aaatactcct tctgggttca accaccttga tgattgtatt 1561 ttgaatactc aggaagtcga aaaggtacac aaaaatactt ttggttgtgc tggagaaagg 1621 agcaagccta aacgtcagaa atccagtact aaactttctg agcttcatga caatcaggac 1681 ggtcttgtga atatggaaag tctcaattcc acacgatctc atgagagaac tggacctgat 1741 gattttgaat ggatgtctga tgaaaggaaa ggaaatgaaa aagatggtgg acacactcag 1801 cattttgaga gccccacaat gaagatccag gagcatccca gcctatctga caccaaacag 1861 cagagaaatc aagatgccgg tgaccaggag gagagctttg tctccgaagt gccccagtcg 1921 gacctgactg cattgtgtga tgaaaagaac tgggaagagc ctatccctgc tttctcctcc 1981 tggcagcggg agaacagtga ctctgatgaa gcccacctct cgccgcaggc tgggcgcctg 2041 atccgtcagc tgctggacga agacagcgac cccatgctct ctcctcggtt ctacgcttat 2101 gggcagagca ggcaatacct ggatgacaca gaagtgcctc cttccccacc aaactcccat 2161 tctttcatga ggcggcgaag ctcctctctg gggtcctatg atgatgagca agaggacctg 2221 acacctgccc agctcacacg aaggattcag agccttaaaa agaagatccg gaagtttgaa 2281 gatagattcg aagaagagaa gaagtacaga ccttcccaca gtgacaaagc agccaatccg 2341 gaggttctga aatggacaaa tgaccttgcc aaattccgga gacaacttaa agaatcaaaa 2401 ctaaagatat ctgaagagga cctaactccc aggatgcggc agcgaagcaa cacactcccc 2461 aagagttttg gttcccaact tgagaaagaa gatgagaaga agcaagagct ggtggataaa 2521 gcaataaagc ccagtgttga agccacattg gaatctattc agaggaagct ccaggagaag 2581 cgagcggaaa gcagccgccc tgaggacatt aaggatatga ccaaagacca gattgctaat 2641 gagaaagtgg ctctgcagaa agctctgtta tattatgaaa gcattcatgg acggccggta 2701 acaaagaacg aacggcaggt gatgaagcca ctatacgaca ggtaccggct ggtcaaacag 2761 atcctctccc gagctaacac catacccatc attggttccc cctccagcaa gcggagaagc 2821 cctttgctgc agccaattat cgagggcgaa actgcttcct tcttcaagga gataaaggaa 2881 gaagaggagg ggtcagaaga cgatagcaat gtgaagccag acttcatggt cactctgaaa 2941 accgatttca gtgcacgatg ctttctggac caattcgaag atgacgctga tggatttatt 3001 tccccaatgg atgataaaat accatcaaaa tgcagccagg acacagggct ttcaaatctc 3061 catgctgcct caatacctga actcctggaa cacctccagg aaatgagaga agaaaagaaa 3121 aggattcgaa agaaacttcg ggattttgaa gacaactttt tcagacagaa tggaagaaat 3181 gtccagaagg aagaccgcac tcctatggct gaagaataca gtgaatataa gcacataaag 3241 gcgaaactga ggctcctgga ggtgctcatc agcaagagag acactgattc caagtccatg 3301 tgaggggcat ggccaagcac agggggctgg cagctgcggt gagagtttac tgtccccaga 3361 gaaagtgcag ctctggaagg cagccttggg gctggccctg caaagcatgc agcccttctg 3421 cctctagacc atttggcatc ggctcctgtt tccattgcct gccttagaaa ctggctggaa 3481 gaagacaatg tgacctgact taggcatttt gtaattggaa agtcaagact gcagtatgtg 3541 cacatgcgca cgcgcatgca cgcacacaca cacacagtag tggagctttc ctaacactag 3601 cagagattaa tcactacatt agacaacact catctacaga gaatatacac tgttcttccc 3661 tggataactg agaaacaaga gaccattctc tgtctaactg tgataaaaac aagctcagga 3721 ctttattcta tagagcaaac ttgctgtgga gggccatgct ctccttggac ccagttaact 3781 gcaaacgtgc attggagccc tatttgctgc cgctgccatt ctagtgacct ttccacagag 3841 ctgcgccttc ctcacgtgtg tgaaaggttt tccccttcag ccctcaggta gatggaagct 3901 gcatctgccc acgatggcag tgcagtcatc atcttcagga tgtttcttca ggacttcctc 3961 agctgacaag gaattttggt ccctgcctag gaccgggtca tctgcagagg acagagagat 4021 ggtaagcagc tgtatgaatg ctgattttaa aaccaggtca tgggagaaga gcctggagat 4081 tctttcctga acactgactg cacttaccag tctgatttta tcgtcaaaca ccaagccagg 4141 ctagcatgct catggcaatc tgtttggggc tgttttgttg tggcactagc caaacataaa 4201 ggggcttaag tcagcctgca tacagaggat cggggagaga aggggcctgt gttctcagcc 4261 tcctgagtac ttaccagagt ttaatttttt taaaaaaaat ctgcactaaa atccccaaac 4321 tgacaggtaa atgtagccct cagagctcag cccaaggcag aatctaaatc acactatttt 4381 cgagatcatg tataaaaaga aaaaaaagaa gtcatgctgt gtggccaatt ataatttttt 4441 tcaaagactt tgtcacaaaa ctgtctatat tagacatttt ggagggacca ggaaatgtaa 4501 gacaccaaat cctccatctc ttcagtgtgc ctgatgtcac ctcatgattt gctgttactt 4561 ttttaactcc tgcgccaagg acagtgggtt ctgtgtccac ctttgtgctt tgcgaggccg 4621 agcccaggca tctgctcgcc tgccacggct gaccagagaa ggtgcttcag gagctctgcc 4681 ttagacgacg tgttacagta tgaacacaca gcagaggcac cctcgtatgt tttgaaagtt 4741 gccttctgaa agggcacagt tttaaggaaa agaaaaagaa tgtaaaacta tactgacccg 4801 ttttcagttt taaagggtcg tgagaaactg gctggtccaa tgggatttac agcaacattt 4861 tccattgctg aagtgaggta gcagctctct tctgtcagct gaatgttaag gatggggaaa 4921 aagaatgcct ttaagtttgc tcttaatcgt atggaagctt gagctatgtg ttggaagtgc 4981 cctggtttta atccatacac aaagacggta cataatccta caggtttaaa tgtacataaa 5041 aatatagttt ggaattcttt gctctactgt ttacattgca gattgctata atttcaagga 5101 gtgagattat aaataaaatg atgcacttta ggatgtttcc tatttttgaa atctgaacat 5161 gaatcattca catgaccaaa aattgtgttt ttttaaaaat acatgtctag tctgtccttt 5221 aatagctctc ttaaataagc tatgatatta atcagatcat taccagttag cttttaaagc 5281 acatttgttt aagactatgt ttttggaaaa atacgctaca gaattttttt ttaagctaca 5341 aataaatgag atgctactaa ttgttttgga atctgttgtt tctgccaaag gtaaattaac 5401 taaagattta ttcaggaatc cccatttgaa tttgtatgat tcaataaaag aaaacaccaa 5461 gtaagttata taaaataaat tgtgtatgag atgttgtgtt ttcctttgta atttccacta 5521 actaactaac taacttatat tcttcatgga atggagccca gaagaaatga gaggaagccc 5581 ttttcacact agatcttatt tgaagaaatg tttgttagtc agtcagtcag tggtttctgg 5641 ctctgccgag ggagatgtgt tccccagcaa ccatttctgc agcccagaat ctcaaggcac 5701 tagaggcggt gtcttaatta attggcttca caaagacaaa atgctctgga ctgggatttt 5761 tcctttgctg tgttgggaat atgtgtttat taattagcac atgccaacaa aataaatgtc 5821 aagagttatt tcataagtgt aagtaaactt aagaattaaa gagtgcagac ttataatttt 5881 ca 

In some embodiments of the methods of the disclosure, the wild type human FAM13A gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_055698.2, transcript variant 1):

(SEQ ID NO: 37) 1 mgagalaicq skaavrlked mkkivavpln eqkdftyqkl fgvslqeler qgltengipa 61 vvwniveylt qhgltqeglf rvngnvkvve qlrlkfesgv pvelgkdgdv csaasllklf 121 lrelpdslit salqprfiql fqdgrndvqe sslrdlikel pdthycllky lcqfltkvak 181 hhvqnrmnvh nlatvfgpnc fhvppglegm keqdlcnkim akilenyntl feveytendh 241 lrcenlarli ivkevyykns lpilltrgle rdmpkpppkt kipksrsegs iqahrvlqpe 301 lsdgipqlsl rlsyrkacle dmnsaegais aklvpssqed erplspfyls ahvpqvsnvs 361 atgellerti rsaveqhlfd vnnsggqsse dsesgtlsas satsarqrrr qskeqdevrh 421 grdkglinke ntpsgfnhld dcilntqeve kvhkntfgca gerskpkrqk sstklselhd 481 nqdglvnmes lnstrshert gpddfewmsd erkgnekdgg htqhfesptm kigehpslsd 541 tkqqrnqdag dqeesfvsev pqsdltalcd eknweepipa fsswqrensd sdeahlspqa 601 grlirqllde dsdpmlsprf yaygqsrqyl ddtevppspp nshsfmrrrs sslgsyddeq 661 edltpaqltr riqslkkkir kfedrfeeek kyrpshsdka anpevlkwtn dlakfrrqlk 721 esklkiseed ltprmrqrsn tlpksfgsql ekedekkqel vdkaikpsve atlesiqrkl 781 qekraessrp edikdmtkdq ianekvalqk allyyesihg rpvtknerqv mkplydryrl 841 vkqilsrant ipiigspssk rrspllqpii egetasffke ikeeeegsed dsnvkpdfmv 901 tlktdfsarc fldqfeddad gfispmddki pskcsqdtgl snlhaasipe llehlqemre 961 ekkrirkklr dfednffrqn grnvqkedrt pmaeeyseyk hikaklrlle vliskrdtds 1021 ksm

In some embodiments of the methods of the disclosure, the wild type human FAM13A gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_001015045.2, transcript variant 2):

(SEQ ID NO: 17) 1 attgaggagc agaaggagta gggtgcgggg gaggaggagg agcgccttta gtgctgcagc 61 agctgctgct ctgattggcc cggtggttca gctgcttccc tggaacaaaa ggtcaaagtg 121 gactgcagtg taaatgtaga gaagcagccg ataaaatagc attgcctgaa gaagtttgga 181 ggctgagagc agcagtagac tggccaactg cagagcaagt tgtttctcca gccgtgcggt 241 gcagcctcat gcccccaacc cagcttagcc actgtaagaa gacgttcact gtacagacga 301 ccaaacttgc cgtggaagag acagttgtga gattcccttg caaatttaca tacgagaatg 361 gcttgtgaaa tcatgcctct gcaaagttca caggaagatg aaagacctct gtcacctttc 421 tatttgagtg ctcatgtacc ccaagtcagc aatgtgtctg caaccggaga actcttagaa 481 agaaccatcc gatcagctgt agaacaacat ctttttgatg ttaataactc tggaggtcaa 541 agttcagagg actcagaatc tggaacacta tcagcatctt ctgccacatc tgccagacag 601 cgccgccgcc agtccaagga gcaggatgaa gttcgacatg ggagagacaa gggacttatc 661 aacaaagaaa atactccttc tgggttcaac caccttgatg attgtatttt gaatactcag 721 gaagtcgaaa aggtacacaa aaatactttt ggttgtgctg gagaaaggag caagcctaaa 781 cgtcagaaat ccagtactaa actttctgag cttcatgaca atcaggacgg tcttgtgaat 841 atggaaagtc tcaattccac acgatctcat gagagaactg gacctgatga ttttgaatgg 901 atgtctgatg aaaggaaagg aaatgaaaaa gatggtggac acactcagca ttttgagagc 961 cccacaatga agatccagga gcatcccagc ctatctgaca ccaaacagca gagaaatcaa 1021 gatgccggtg accaggagga gagctttgtc tccgaagtgc cccagtcgga cctgactgca 1081 ttgtgtgatg aaaagaactg ggaagagcct atccctgctt tctcctcctg gcagcgggag 1141 aacagtgact ctgatgaagc ccacctctcg ccgcaggctg ggcgcctgat ccgtcagctg 1201 ctggacgaag acagcgaccc catgctctct cctcggttct acgcttatgg gcagagcagg 1261 caatacctgg atgacacaga agtgcctcct tccccaccaa actcccattc tttcatgagg 1321 cggcgaagct cctctctggg gtcctatgat gatgagcaag aggacctgac acctgcccag 1381 ctcacacgaa ggattcagag ccttaaaaag aagatccgga agtttgaaga tagattcgaa 1441 gaagagaaga agtacagacc ttcccacagt gacaaagcag ccaatccgga ggttctgaaa 1501 tggacaaatg accttgccaa attccggaga caacttaaag aatcaaaact aaagatatct 1561 gaagaggacc taactcccag gatgcggcag cgaagcaaca cactccccaa gagttttggt 1621 tcccaacttg agaaagaaga tgagaagaag caagagctgg tggataaagc aataaagccc 1681 agtgttgaag ccacattgga atctattcag aggaagctcc aggagaagcg agcggaaagc 1741 agccgccctg aggacattaa ggatatgacc aaagaccaga ttgctaatga gaaagtggct 1801 ctgcagaaag ctctgttata ttatgaaagc attcatggac ggccggtaac aaagaacgaa 1861 cggcaggtga tgaagccact atacgacagg taccggctgg tcaaacagat cctctcccga 1921 gctaacacca tacccatcat tggttccccc tccagcaagc ggagaagccc tttgctgcag 1981 ccaattatcg agggcgaaac tgcttccttc ttcaaggaga taaaggaaga agaggagggg 2041 tcagaagacg atagcaatgt gaagccagac ttcatggtca ctctgaaaac cgatttcagt 2101 gcacgatgct ttctggacca attcgaagat gacgctgatg gatttatttc cccaatggat 2161 gataaaatac catcaaaatg cagccaggac acagggcttt caaatctcca tgctgcctca 2221 atacctgaac tcctggaaca cctccaggaa atgagagaag aaaagaaaag gattcgaaag 2281 aaacttcggg attttgaaga caactttttc agacagaatg gaagaaatgt ccagaaggaa 2341 gaccgcactc ctatggctga agaatacagt gaatataagc acataaaggc gaaactgagg 2401 ctcctggagg tgctcatcag caagagagac actgattcca agtccatgtg aggggcatgg 2461 ccaagcacag ggggctggca gctgcggtga gagtttactg tccccagaga aagtgcagct 2521 ctggaaggca gccttggggc tggccctgca aagcatgcag cccttctgcc tctagaccat 2581 ttggcatcgg ctcctgtttc cattgcctgc cttagaaact ggctggaaga agacaatgtg 2641 acctgactta ggcattttgt aattggaaag tcaagactgc agtatgtgca catgcgcacg 2701 cgcatgcacg cacacacaca cacagtagtg gagctttcct aacactagca gagattaatc 2761 actacattag acaacactca tctacagaga atatacactg ttcttccctg gataactgag 2821 aaacaagaga ccattctctg tctaactgtg ataaaaacaa gctcaggact ttattctata 2881 gagcaaactt gctgtggagg gccatgctct ccttggaccc agttaactgc aaacgtgcat 2941 tggagcccta tttgctgccg ctgccattct agtgaccttt ccacagagct gcgccttcct 3001 cacgtgtgtg aaaggttttc cccttcagcc ctcaggtaga tggaagctgc atctgcccac 3061 gatggcagtg cagtcatcat cttcaggatg tttcttcagg acttcctcag ctgacaagga 3121 attttggtcc ctgcctagga ccgggtcatc tgcagaggac agagagatgg taagcagctg 3181 tatgaatgct gattttaaaa ccaggtcatg ggagaagagc ctggagattc tttcctgaac 3241 actgactgca cttaccagtc tgattttatc gtcaaacacc aagccaggct agcatgctca 3301 tggcaatctg tttggggctg ttttgttgtg gcactagcca aacataaagg ggcttaagtc 3361 agcctgcata cagaggatcg gggagagaag gggcctgtgt tctcagcctc ctgagtactt 3421 accagagttt aattttttta aaaaaaatct gcactaaaat ccccaaactg acaggtaaat 3481 gtagccctca gagctcagcc caaggcagaa tctaaatcac actattttcg agatcatgta 3541 taaaaagaaa aaaaagaagt catgctgtgt ggccaattat aatttttttc aaagactttg 3601 tcacaaaact gtctatatta gacattttgg agggaccagg aaatgtaaga caccaaatcc 3661 tccatctctt cagtgtgcct gatgtcacct catgatttgc tgttactttt ttaactcctg 3721 cgccaaggac agtgggttct gtgtccacct ttgtgctttg cgaggccgag cccaggcatc 3781 tgctcgcctg ccacggctga ccagagaagg tgcttcagga gctctgcctt agacgacgtg 3841 ttacagtatg aacacacagc agaggcaccc tcgtatgttt tgaaagttgc cttctgaaag 3901 ggcacagttt taaggaaaag aaaaagaatg taaaactata ctgacccgtt ttcagtttta 3961 aagggtcgtg agaaactggc tggtccaatg ggatttacag caacattttc cattgctgaa 4021 gtgaggtagc agctctcttc tgtcagctga atgttaagga tggggaaaaa gaatgccttt 4081 aagtttgctc ttaatcgtat ggaagcttga gctatgtgtt ggaagtgccc tggttttaat 4141 ccatacacaa agacggtaca taatcctaca ggtttaaatg tacataaaaa tatagtttgg 4201 aattctttgc tctactgttt acattgcaga ttgctataat ttcaaggagt gagattataa 4261 ataaaatgat gcactttagg atgtttccta tttttgaaat ctgaacatga atcattcaca 4321 tgaccaaaaa ttgtgttttt ttaaaaatac atgtctagtc tgtcctttaa tagctctctt 4381 aaataagcta tgatattaat cagatcatta ccagttagct tttaaagcac atttgtttaa 4441 gactatgttt ttggaaaaat acgctacaga attttttttt aagctacaaa taaatgagat 4501 gctactaatt gttttggaat ctgttgtttc tgccaaaggt aaattaacta aagatttatt 4561 caggaatccc catttgaatt tgtatgattc aataaaagaa aacaccaagt aagttatata 4621 aaataaattg tgtatgagat gttgtgtttt cctttgtaat ttccactaac taactaacta 4681 acttatattc ttcatggaat ggagcccaga agaaatgaga ggaagccctt ttcacactag 4741 atcttatttg aagaaatgtt tgttagtcag tcagtcagtg gtttctggct ctgccgaggg 4801 agatgtgttc cccagcaacc atttctgcag cccagaatct caaggcacta gaggcggtgt 4861 cttaattaat tggcttcaca aagacaaaat gctctggact gggatttttc ctttgctgtg 4921 ttgggaatat gtgtttatta attagcacat gccaacaaaa taaatgtcaa gagttatttc 4981 ataagtgtaa gtaaacttaa gaattaaaga gtgcagactt ataattttca 

In some embodiments of the methods of the disclosure, the wild type human FAM13A gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_001015045.1, transcript variant 2):

(SEQ ID NO: 18) 1 maceimplqs sqederplsp fylsahvpqv snvsatgell ertirsaveq hlfdvnnsgg 61 qssedsesgt lsassatsar qrrrqskeqd evrhgrdkgl inkentpsgf nhlddcilnt 121 qevekvhknt fgcagerskp krqksstkls elhdnqdglv nmeslnstrs hertgpddfe 181 wmsderkgne kdgghtqhfe sptmkigehp slsdtkqqrn qdagdqeesf vsevpqsdlt 241 alcdeknwee pipafsswqr ensdsdeahl spqagrlirq lldedsdpml sprfyaygqs 301 rqylddtevp psppnshsfm rrrssslgsy ddeqedltpa qltrriqslk kkirkfedrf 361 eeekkyrpsh sdkaanpevl kwtndlakfr rqlkesklki seedltprmr qrsntlpksf 421 gsqlekedek kqelvdkaik psveatlesi qrklqekrae ssrpedikdm tkdqianekv 481 alqkallyye sihgrpvtkn erqvmkplyd ryrlvkqils rantipiigs psskrrspll 541 qpiiegetas ffkeikeeee gseddsnvkp dfmvtlktdf sarcfldqfe ddadgfispm 601 ddkipskcsq dtglsnlhaa sipellehlq emreekkrir kklrdfednf frqngrnvqk 661 edrtpmaeey seykhikakl rllevliskr dtdsksm

In some embodiments of the methods of the disclosure, the wild type human FAM13A gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_001265578.1, transcript variant 3):

(SEQ ID NO: 38) 1 attgaggagc agaaggagta gggtgcgggg gaggaggagg agcgccttta gtgctgcagc 61 agctgctgct ctgattggcc cggtggttca gctgcttccc tggaacaaaa ggtcaaagtg 121 gactgcagtg taaatgtaga gaagcagccg ataaaatagc attgcctgaa gaagtttgga 181 ggctgagagc agcagtagac tggccaactg cagagcaagt tgtttctcca gccgtgcggt 241 gcagcctcat gcccccaacc cagcttagcc actgtaagaa gacgttcact gtacagacga 301 ccaaacttgc cgtggaagag acagttgtga gattcccttg caaatttaca tacgagaatg 361 gcttgtgaaa tcatgcctct gcaaagtgct catgtacccc aagtcagcaa tgtgtctgca 421 accggagaac tcttagaaag aaccatccga tcagctgtag aacaacatct ttttgatgtt 481 aataactctg gaggtcaaag ttcagaggac tcagaatctg gaacactatc agcatcttct 541 gccacatctg ccagacagcg ccgccgccag tccaaggagc aggatgaagt tcgacatggg 601 agagacaagg gacttatcaa caaagaaaat actccttctg ggttcaacca ccttgatgat 661 tgtattttga atactcagga agtcgaaaag gtacacaaaa atacttttgg ttgtgctgga 721 gaaaggagca agcctaaacg tcagaaatcc agtactaaac tttctgagct tcatgacaat 781 caggacggtc ttgtgaatat ggaaagtctc aattccacac gatctcatga gagaactgga 841 cctgatgatt ttgaatggat gtctgatgaa aggaaaggaa atgaaaaaga tggtggacac 901 actcagcatt ttgagagccc cacaatgaag atccaggagc atcccagcct atctgacacc 961 aaacagcaga gaaatcaaga tgccggtgac caggaggaga gctttgtctc cgaagtgccc 1021 cagtcggacc tgactgcatt gtgtgatgaa aagaactggg aagagcctat ccctgctttc 1081 tcctcctggc agcgggagaa cagtgactct gatgaagccc acctctcgcc gcaggctggg 1141 cgcctgatcc gtcagctgct ggacgaagac agcgacccca tgctctctcc tcggttctac 1201 gcttatgggc agagcaggca atacctggat gacacagaag tgcctccttc cccaccaaac 1261 tcccattctt tcatgaggcg gcgaagctcc tctctggggt cctatgatga tgagcaagag 1321 gacctgacac ctgcccagct cacacgaagg attcagagcc ttaaaaagaa gatccggaag 1381 tttgaagata gattcgaaga agagaagaag tacagacctt cccacagtga caaagcagcc 1441 aatccggagg ttctgaaatg gacaaatgac cttgccaaat tccggagaca acttaaagaa 1501 tcaaaactaa agatatctga agaggaccta actcccagga tgcggcagcg aagcaacaca 1561 ctccccaaga gttttggttc ccaacttgag aaagaagatg agaagaagca agagctggtg 1621 gataaagcaa taaagcccag tgttgaagcc acattggaat ctattcagag gaagctccag 1681 gagaagcgag cggaaagcag ccgccctgag gacattaagg atatgaccaa agaccagatt 1741 gctaatgaga aagtggctct gcagaaagct ctgttatatt atgaaagcat tcatggacgg 1801 ccggtaacaa agaacgaacg gcaggtgatg aagccactat acgacaggta ccggctggtc 1861 aaacagatcc tctcccgagc taacaccata cccatcattg gttccccctc cagcaagcgg 1921 agaagccctt tgctgcagcc aattatcgag ggcgaaactg cttccttctt caaggagata 1981 aaggaagaag aggaggggtc agaagacgat agcaatgtga agccagactt catggtcact 2041 ctgaaaaccg atttcagtgc acgatgcttt ctggaccaat tcgaagatga cgctgatgga 2101 tttatttccc caatggatga taaaatacca tcaaaatgca gccaggacac agggctttca 2161 aatctccatg ctgcctcaat acctgaactc ctggaacacc tccaggaaat gagagaagaa 2221 aagaaaagga ttcgaaagaa acttcgggat tttgaagaca actttttcag acagaatgga 2281 agaaatgtcc agaaggaaga ccgcactcct atggctgaag aatacagtga atataagcac 2341 ataaaggcga aactgaggct cctggaggtg ctcatcagca agagagacac tgattccaag 2401 tccatgtgag gggcatggcc aagcacaggg ggctggcagc tgcggtgaga gtttactgtc 2461 cccagagaaa gtgcagctct ggaaggcagc cttggggctg gccctgcaaa gcatgcagcc 2521 cttctgcctc tagaccattt ggcatcggct cctgtttcca ttgcctgcct tagaaactgg 2581 ctggaagaag acaatgtgac ctgacttagg cattttgtaa ttggaaagtc aagactgcag 2641 tatgtgcaca tgcgcacgcg catgcacgca cacacacaca cagtagtgga gctttcctaa 2701 cactagcaga gattaatcac tacattagac aacactcatc tacagagaat atacactgtt 2761 cttccctgga taactgagaa acaagagacc attctctgtc taactgtgat aaaaacaagc 2821 tcaggacttt attctataga gcaaacttgc tgtggagggc catgctctcc ttggacccag 2881 ttaactgcaa acgtgcattg gagccctatt tgctgccgct gccattctag tgacctttcc 2941 acagagctgc gccttcctca cgtgtgtgaa aggttttccc cttcagccct caggtagatg 3001 gaagctgcat ctgcccacga tggcagtgca gtcatcatct tcaggatgtt tcttcaggac 3061 ttcctcagct gacaaggaat tttggtccct gcctaggacc gggtcatctg cagaggacag 3121 agagatggta agcagctgta tgaatgctga ttttaaaacc aggtcatggg agaagagcct 3181 ggagattctt tcctgaacac tgactgcact taccagtctg attttatcgt caaacaccaa 3241 gccaggctag catgctcatg gcaatctgtt tggggctgtt ttgttgtggc actagccaaa 3301 cataaagggg cttaagtcag cctgcataca gaggatcggg gagagaaggg gcctgtgttc 3361 tcagcctcct gagtacttac cagagtttaa tttttttaaa aaaaatctgc actaaaatcc 3421 ccaaactgac aggtaaatgt agccctcaga gctcagccca aggcagaatc taaatcacac 3481 tattttcgag atcatgtata aaaagaaaaa aaagaagtca tgctgtgtgg ccaattataa 3541 tttttttcaa agactttgtc acaaaactgt ctatattaga cattttggag ggaccaggaa 3601 atgtaagaca ccaaatcctc catctcttca gtgtgcctga tgtcacctca tgatttgctg 3661 ttactttttt aactcctgcg ccaaggacag tgggttctgt gtccaccttt gtgctttgcg 3721 aggccgagcc caggcatctg ctcgcctgcc acggctgacc agagaaggtg cttcaggagc 3781 tctgccttag acgacgtgtt acagtatgaa cacacagcag aggcaccctc gtatgttttg 3841 aaagttgcct tctgaaaggg cacagtttta aggaaaagaa aaagaatgta aaactatact 3901 gacccgtttt cagttttaaa gggtcgtgag aaactggctg gtccaatggg atttacagca 3961 acattttcca ttgctgaagt gaggtagcag ctctcttctg tcagctgaat gttaaggatg 4021 gggaaaaaga atgcctttaa gtttgctctt aatcgtatgg aagcttgagc tatgtgttgg 4081 aagtgccctg gttttaatcc atacacaaag acggtacata atcctacagg tttaaatgta 4141 cataaaaata tagtttggaa ttctttgctc tactgtttac attgcagatt gctataattt 4201 caaggagtga gattataaat aaaatgatgc actttaggat gtttcctatt tttgaaatct 4261 gaacatgaat cattcacatg accaaaaatt gtgttttttt aaaaatacat gtctagtctg 4321 tcctttaata gctctcttaa ataagctatg atattaatca gatcattacc agttagcttt 4381 taaagcacat ttgtttaaga ctatgttttt ggaaaaatac gctacagaat ttttttttaa 4441 gctacaaata aatgagatgc tactaattgt tttggaatct gttgtttctg ccaaaggtaa 4501 attaactaaa gatttattca ggaatcccca tttgaatttg tatgattcaa taaaagaaaa 4561 caccaagtaa gttatataaa ataaattgtg tatgagatgt tgtgttttcc tttgtaattt 4621 ccactaacta actaactaac ttatattctt catggaatgg agcccagaag aaatgagagg 4681 aagccctttt cacactagat cttatttgaa gaaatgtttg ttagtcagtc agtcagtggt 4741 ttctggctct gccgagggag atgtgttccc cagcaaccat ttctgcagcc cagaatctca 4801 aggcactaga ggcggtgtct taattaattg gcttcacaaa gacaaaatgc tctggactgg 4861 gatttttcct ttgctgtgtt gggaatatgt gtttattaat tagcacatgc caacaaaata 4921 aatgtcaaga gttatttcat aagtgtaagt aaacttaaga attaaagagt gcagacttat 4981 aattttca

In some embodiments of the methods of the disclosure, the wild type human FAM13A gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_001252507.1, transcript variant 3):

(SEQ ID NO: 39) 1 maceimplqs ahvpqvsnvs atgellerti rsaveqhlfd vnnsggqsse dsesgtlsas 61 satsarqrrr qskeqdevrh grdkglinke ntpsgfnhld dcilntqeve kvhkntfgca 121 gerskpkrqk sstklselhd nqdglvnmes lnstrshert gpddfewmsd erkgnekdgg 181 htqhfesptm kigehpslsd tkqqrnqdag dqeesfvsev pqsdltalcd eknweepipa 241 fsswqrensd sdeahlspqa grlirqllde dsdpmlsprf yayggsrqyl ddtevppspp 301 nshsfmrrrs sslgsyddeq edltpaqltr riqslkkkir kfedrfeeek kyrpshsdka 361 anpevlkwtn dlakfrrqlk esklkiseed ltprmrqrsn tlpksfgsql ekedekkqel 421 vdkaikpsve atlesiqrkl qekraessrp edikdmtkdq ianekvalqk allyyesihg 481 rpvtknerqv mkplydryrl vkqilsrant ipiigspssk rrspllqpii egetasffke 541 ikeeeegsed dsnvkpdfmv tlktdfsarc fldqfeddad gfispmddki pskcsqdtgl 601 snlhaasipe llehlqemre ekkrirkklr dfednffrqn grnvqkedrt pmaeeyseyk 661 hikaklrlle vliskrdtds ksm

In some embodiments of the methods of the disclosure, the wild type human FAM13A gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_001265579.1, transcript variant 4):

(SEQ ID NO: 40) 1 attgaggagc agaaggagta gggtgcgggg gaggaggagg agcgccttta gtgctgcagc 61 agctgctgct ctgattggcc cggtggttca gctgcttccc tggaacaaaa ggtcaaagtg 121 gactgcagtg taaatgtaga gaagcagccg ataaaatagc attgcctgaa gaagtttgga 181 ggctgagagc agcagtagac tggccaactg cagagcaagt tgtttctcca gccgtgcggt 241 gcagcctcat gcccccaacc cagcttagcc actgtaagaa gacgttcact gtacagacga 301 ccaaacttgc cgtggaagag acagttgtga gattcccttg caaatttaca tacgagaatg 361 gcttgtgaaa tcatgcctct gcaaagttca caggaagatg aaagacctct gtcacctttc 421 tatttgagtg ctcatgtacc ccaagtcagc aatgtgtctg caaccggaga actcttagaa 481 agaaccatcc gatcagctgt agaacaacat ctttttgatg ttaataactc tggaggtcaa 541 agttcagagg actcagaatc tggaacacta tcagcatctt ctgccacatc tgccagacag 601 cgccgccgcc agtccaagga gcaggatgaa gttcgacatg ggagagacaa gggacttatc 661 aacaaagaaa atactccttc tgggttcaac caccttgatg attgtatttt gaatactcag 721 gaagtcgaaa aggtacacaa aaatactttt ggttgtgctg gagaaaggag caagcctaaa 781 cgtcagaaat ccagtactaa actttctgag cttcatgaca atcaggacgg tcttgtgaat 841 atggaaagtc tcaattccac acgatctcat gagagaactg gacctgatga ttttgaatgg 901 atgtctgatg aaaggaaagg aaatgaaaaa gatggtggac acactcagca ttttgagagc 961 cccacaatga agatccagga gcatcccagc ctatctgaca ccaaacagca gagaaatcaa 1021 gatgccggtg accaggagga gagctttgtc tccgaagtgc cccagtcgga cctgactgca 1081 ttgtgtgatg aaaagaactg ggaagagcct atccctgctt tctcctcctg gcagcgggag 1141 aacagtgact ctgatgaagc ccacctctcg ccgcaggctg ggcgcctgat ccgtcagctg 1201 ctggacgaag acagcgaccc catgctctct cctcggttct acgcttatgg gcagagcagg 1261 caatacctgg atgacacaga agtgcctcct tccccaccaa actcccattc tttcatgagg 1321 cggcgaagct cctctctggg gtcctatgat gatgagcaag aggacctgac acctgcccag 1381 ctcacacgaa ggattcagag ccttaaaaag aagatccgga agtttgaaga tagattcgaa 1441 gaagagaaga agtacagacc ttcccacagt gacaaagcag ccaatccgga ggttctgaaa 1501 tggacaaatg accttgccaa attccggaga caacttaaag aatcaaaact aaagatatct 1561 gaagaggacc taactcccag gatgcggcag cgaagcaaca cactccccaa gagttttggt 1621 tcccaacttg agaaagaaga tgagaagaag caagagctgg tggataaagc aataaagccc 1681 agtgttgaag ccacattgga atctattcag aggaagctcc aggagaagcg agcggaaagc 1741 agccgccctg aggacattaa ggatatgacc aaagaccaga ttgctaatga gaaagtggct 1801 ctgcagaaag ctctgttata ttatgaaagc attcatggac ggccggtaac aaagaacgaa 1861 cggcaggtga tgaagccact atacgacagg taccggctgg tcaaacagat cctctcccga 1921 gctaacacca tacccatcat tgaagaagag gaggggtcag aagacgatag caatgtgaag 1981 ccagacttca tggtcactct gaaaaccgat ttcagtgcac gatgctttct ggaccaattc 2041 gaagatgacg ctgatggatt tatttcccca atggatgata aaataccatc aaaatgcagc 2101 caggacacag ggctttcaaa tctccatgct gcctcaatac ctgaactcct ggaacacctc 2161 caggaaatga gagaagaaaa gaaaaggatt cgaaagaaac ttcgggattt tgaagacaac 2221 tttttcagac agaatggaag aaatgtccag aaggaagacc gcactcctat ggctgaagaa 2281 tacagtgaat ataagcacat aaaggcgaaa ctgaggctcc tggaggtgct catcagcaag 2341 agagacactg attccaagtc catgtgaggg gcatggccaa gcacaggggg ctggcagctg 2401 cggtgagagt ttactgtccc cagagaaagt gcagctctgg aaggcagcct tggggctggc 2461 cctgcaaagc atgcagccct tctgcctcta gaccatttgg catcggctcc tgtttccatt 2521 gcctgcctta gaaactggct ggaagaagac aatgtgacct gacttaggca ttttgtaatt 2581 ggaaagtcaa gactgcagta tgtgcacatg cgcacgcgca tgcacgcaca cacacacaca 2641 gtagtggagc tttcctaaca ctagcagaga ttaatcacta cattagacaa cactcatcta 2701 cagagaatat acactgttct tccctggata actgagaaac aagagaccat tctctgtcta 2761 actgtgataa aaacaagctc aggactttat tctatagagc aaacttgctg tggagggcca 2821 tgctctcctt ggacccagtt aactgcaaac gtgcattgga gccctatttg ctgccgctgc 2881 cattctagtg acctttccac agagctgcgc cttcctcacg tgtgtgaaag gttttcccct 2941 tcagccctca ggtagatgga agctgcatct gcccacgatg gcagtgcagt catcatcttc 3001 aggatgtttc ttcaggactt cctcagctga caaggaattt tggtccctgc ctaggaccgg 3061 gtcatctgca gaggacagag agatggtaag cagctgtatg aatgctgatt ttaaaaccag 3121 gtcatgggag aagagcctgg agattctttc ctgaacactg actgcactta ccagtctgat 3181 tttatcgtca aacaccaagc caggctagca tgctcatggc aatctgtttg gggctgtttt 3241 gttgtggcac tagccaaaca taaaggggct taagtcagcc tgcatacaga ggatcgggga 3301 gagaaggggc ctgtgttctc agcctcctga gtacttacca gagtttaatt tttttaaaaa 3361 aaatctgcac taaaatcccc aaactgacag gtaaatgtag ccctcagagc tcagcccaag 3421 gcagaatcta aatcacacta ttttcgagat catgtataaa aagaaaaaaa agaagtcatg 3481 ctgtgtggcc aattataatt tttttcaaag actttgtcac aaaactgtct atattagaca 3541 ttttggaggg accaggaaat gtaagacacc aaatcctcca tctcttcagt gtgcctgatg 3601 tcacctcatg atttgctgtt acttttttaa ctcctgcgcc aaggacagtg ggttctgtgt 3661 ccacctttgt gctttgcgag gccgagccca ggcatctgct cgcctgccac ggctgaccag 3721 agaaggtgct tcaggagctc tgccttagac gacgtgttac agtatgaaca cacagcagag 3781 gcaccctcgt atgttttgaa agttgccttc tgaaagggca cagttttaag gaaaagaaaa 3841 agaatgtaaa actatactga cccgttttca gttttaaagg gtcgtgagaa actggctggt 3901 ccaatgggat ttacagcaac attttccatt gctgaagtga ggtagcagct ctcttctgtc 3961 agctgaatgt taaggatggg gaaaaagaat gcctttaagt ttgctcttaa tcgtatggaa 4021 gcttgagcta tgtgttggaa gtgccctggt tttaatccat acacaaagac ggtacataat 4081 cctacaggtt taaatgtaca taaaaatata gtttggaatt ctttgctcta ctgtttacat 4141 tgcagattgc tataatttca aggagtgaga ttataaataa aatgatgcac tttaggatgt 4201 ttcctatttt tgaaatctga acatgaatca ttcacatgac caaaaattgt gtttttttaa 4261 aaatacatgt ctagtctgtc ctttaatagc tctcttaaat aagctatgat attaatcaga 4321 tcattaccag ttagctttta aagcacattt gtttaagact atgtttttgg aaaaatacgc 4381 tacagaattt ttttttaagc tacaaataaa tgagatgcta ctaattgttt tggaatctgt 4441 tgtttctgcc aaaggtaaat taactaaaga tttattcagg aatccccatt tgaatttgta 4501 tgattcaata aaagaaaaca ccaagtaagt tatataaaat aaattgtgta tgagatgttg 4561 tgttttcctt tgtaatttcc actaactaac taactaactt atattcttca tggaatggag 4621 cccagaagaa atgagaggaa gcccttttca cactagatct tatttgaaga aatgtttgtt 4681 agtcagtcag tcagtggttt ctggctctgc cgagggagat gtgttcccca gcaaccattt 4741 ctgcagccca gaatctcaag gcactagagg cggtgtctta attaattggc ttcacaaaga 4801 caaaatgctc tggactggga tttttccttt gctgtgttgg gaatatgtgt ttattaatta 4861 gcacatgcca acaaaataaa tgtcaagagt tatttcataa gtgtaagtaa acttaagaat 4921 taaagagtgc agacttataa ttttca 

In some embodiments of the methods of the disclosure, the wild type human FAM13A gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_001252508.1, transcript variant 4):

(SEQ ID NO: 41) 1 maceimplqs sqederplsp fylsahvpqv snvsatgell ertirsaveq hlfdvnnsgg 61 qssedsesgt lsassatsar qrrrqskeqd evrhgrdkgl inkentpsgf nhlddcilnt 121 qevekvhknt fgcagerskp krqksstkls elhdnqdglv nmeslnstrs hertgpddfe 181 wmsderkgne kdgghtqhfe sptmkigehp slsdtkqqrn qdagdqeesf vsevpqsdlt 241 alcdeknwee pipafsswqr ensdsdeahl spqagrlirq lldedsdpml sprfyaygqs 301 rqylddtevp psppnshsfm rrrssslgsy ddeqedltpa qltrriqslk kkirkfedrf 361 eeekkyrpsh sdkaanpevl kwtndlakfr rqlkesklki seedltprmr qrsntlpksf 421 gsqlekedek kqelvdkaik psveatlesi qrklqekrae ssrpedikdm tkdqianekv 481 alqkallyye sihgrpvtkn erqvmkplyd ryrlvkqils rantipiiee eegseddsnv 541 kpdfmvtlkt dfsarcfldq feddadgfis pmddkipskc sqdtglsnlh aasipelleh 601 lqemreekkr irkklrdfed nffrqngrnv qkedrtpmae eyseykhika klrllevlis 661 krdtdsksm

In some embodiments of the methods of the disclosure, the wild type human FAM13A gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_001265580.1, transcript variant 5):

(SEQ ID NO: 42) 1 attgaggagc agaaggagta gggtgcgggg gaggaggagg agcgccttta gtgctgcagc 61 agctgctgct ctgattggcc cggtggttca gctgcttccc tggaacaaaa ggtcaaagtg 121 gactgcagtg taaatgtaga gaagcagccg ataaaatagc attgcctgaa gaagtttgga 181 ggctgagagc agcagtagac tggccaactg cagagcaagt tgtttctcca gccgtgcggt 241 gcagcctcat gcccccaacc cagcttagcc actgtaagaa gacgttcact gtacagacga 301 ccaaacttgc cgtggaagag acagttgtga gattcccttg caaatttaca tacgagaatg 361 gcttgtgaaa tcatgcctct gcaaagactc ttagaaagaa ccatccgatc agctgtagaa 421 caacatcttt ttgatgttaa taactctgga ggtcaaagtt cagaggactc agaatctgga 481 acactatcag catcttctgc cacatctgcc agacagcgcc gccgccagtc caaggagcag 541 gatgaagttc gacatgggag agacaaggga cttatcaaca aagaaaatac tccttctggg 601 ttcaaccacc ttgatgattg tattttgaat actcaggaag tcgaaaaggt acacaaaaat 661 acttttggtt gtgctggaga aaggagcaag cctaaacgtc agaaatccag tactaaactt 721 tctgagcttc atgacaatca ggacggtctt gtgaatatgg aaagtctcaa ttccacacga 781 tctcatgaga gaactggacc tgatgatttt gaatggatgt ctgatgaaag gaaaggaaat 841 gaaaaagatg gtggacacac tcagcatttt gagagcccca caatgaagat ccaggagcat 901 cccagcctat ctgacaccaa acagcagaga aatcaagatg ccggtgacca ggaggagagc 961 tttgtctccg aagtgcccca gtcggacctg actgcattgt gtgatgaaaa gaactgggaa 1021 gagcctatcc ctgctttctc ctcctggcag cgggagaaca gtgactctga tgaagcccac 1081 ctctcgccgc aggctgggcg cctgatccgt cagctgctgg acgaagacag cgaccccatg 1141 ctctctcctc ggttctacgc ttatgggcag agcaggcaat acctggatga cacagaagtg 1201 cctccttccc caccaaactc ccattctttc atgaggcggc gaagctcctc tctggggtcc 1261 tatgatgatg agcaagagga cctgacacct gcccagctca cacgaaggat tcagagcctt 1321 aaaaagaaga tccggaagtt tgaagataga ttcgaagaag agaagaagta cagaccttcc 1381 cacagtgaca aagcagccaa tccggaggtt ctgaaatgga caaatgacct tgccaaattc 1441 cggagacaac ttaaagaatc aaaactaaag atatctgaag aggacctaac tcccaggatg 1501 cggcagcgaa gcaacacact ccccaagagt tttggttccc aacttgagaa agaagatgag 1561 aagaagcaag agctggtgga taaagcaata aagcccagtg ttgaagccac attggaatct 1621 attcagagga agctccagga gaagcgagcg gaaagcagcc gccctgagga cattaaggat 1681 atgaccaaag accagattgc taatgagaaa gtggctctgc agaaagctct gttatattat 1741 gaaagcattc atggacggcc ggtaacaaag aacgaacggc aggtgatgaa gccactatac 1801 gacaggtacc ggctggtcaa acagatcctc tcccgagcta acaccatacc catcattggt 1861 tccccctcca gcaagcggag aagccctttg ctgcagccaa ttatcgaggg cgaaactgct 1921 tccttcttca aggagataaa ggaagaagag gaggggtcag aagacgatag caatgtgaag 1981 ccagacttca tggtcactct gaaaaccgat ttcagtgcac gatgctttct ggaccaattc 2041 gaagatgacg ctgatggatt tatttcccca atggatgata aaataccatc aaaatgcagc 2101 caggacacag ggctttcaaa tctccatgct gcctcaatac ctgaactcct ggaacacctc 2161 caggaaatga gagaagaaaa gaaaaggatt cgaaagaaac ttcgggattt tgaagacaac 2221 tttttcagac agaatggaag aaatgtccag aaggaagacc gcactcctat ggctgaagaa 2281 tacagtgaat ataagcacat aaaggcgaaa ctgaggctcc tggaggtgct catcagcaag 2341 agagacactg attccaagtc catgtgaggg gcatggccaa gcacaggggg ctggcagctg 2401 cggtgagagt ttactgtccc cagagaaagt gcagctctgg aaggcagcct tggggctggc 2461 cctgcaaagc atgcagccct tctgcctcta gaccatttgg catcggctcc tgtttccatt 2521 gcctgcctta gaaactggct ggaagaagac aatgtgacct gacttaggca ttttgtaatt 2581 ggaaagtcaa gactgcagta tgtgcacatg cgcacgcgca tgcacgcaca cacacacaca 2641 gtagtggagc tttcctaaca ctagcagaga ttaatcacta cattagacaa cactcatcta 2701 cagagaatat acactgttct tccctggata actgagaaac aagagaccat tctctgtcta 2761 actgtgataa aaacaagctc aggactttat tctatagagc aaacttgctg tggagggcca 2821 tgctctcctt ggacccagtt aactgcaaac gtgcattgga gccctatttg ctgccgctgc 2881 cattctagtg acctttccac agagctgcgc cttcctcacg tgtgtgaaag gttttcccct 2941 tcagccctca ggtagatgga agctgcatct gcccacgatg gcagtgcagt catcatcttc 3001 aggatgtttc ttcaggactt cctcagctga caaggaattt tggtccctgc ctaggaccgg 3061 gtcatctgca gaggacagag agatggtaag cagctgtatg aatgctgatt ttaaaaccag 3121 gtcatgggag aagagcctgg agattctttc ctgaacactg actgcactta ccagtctgat 3181 tttatcgtca aacaccaagc caggctagca tgctcatggc aatctgtttg gggctgtttt 3241 gttgtggcac tagccaaaca taaaggggct taagtcagcc tgcatacaga ggatcgggga 3301 gagaaggggc ctgtgttctc agcctcctga gtacttacca gagtttaatt tttttaaaaa 3361 aaatctgcac taaaatcccc aaactgacag gtaaatgtag ccctcagagc tcagcccaag 3421 gcagaatcta aatcacacta ttttcgagat catgtataaa aagaaaaaaa agaagtcatg 3481 ctgtgtggcc aattataatt tttttcaaag actttgtcac aaaactgtct atattagaca 3541 ttttggaggg accaggaaat gtaagacacc aaatcctcca tctcttcagt gtgcctgatg 3601 tcacctcatg atttgctgtt acttttttaa ctcctgcgcc aaggacagtg ggttctgtgt 3661 ccacctttgt gctttgcgag gccgagccca ggcatctgct cgcctgccac ggctgaccag 3721 agaaggtgct tcaggagctc tgccttagac gacgtgttac agtatgaaca cacagcagag 3781 gcaccctcgt atgttttgaa agttgccttc tgaaagggca cagttttaag gaaaagaaaa 3841 agaatgtaaa actatactga cccgttttca gttttaaagg gtcgtgagaa actggctggt 3901 ccaatgggat ttacagcaac attttccatt gctgaagtga ggtagcagct ctcttctgtc 3961 agctgaatgt taaggatggg gaaaaagaat gcctttaagt ttgctcttaa tcgtatggaa 4021 gcttgagcta tgtgttggaa gtgccctggt tttaatccat acacaaagac ggtacataat 4081 cctacaggtt taaatgtaca taaaaatata gtttggaatt ctttgctcta ctgtttacat 4141 tgcagattgc tataatttca aggagtgaga ttataaataa aatgatgcac tttaggatgt 4201 ttcctatttt tgaaatctga acatgaatca ttcacatgac caaaaattgt gtttttttaa 4261 aaatacatgt ctagtctgtc ctttaatagc tctcttaaat aagctatgat attaatcaga 4321 tcattaccag ttagctttta aagcacattt gtttaagact atgtttttgg aaaaatacgc 4381 tacagaattt ttttttaagc tacaaataaa tgagatgcta ctaattgttt tggaatctgt 4441 tgtttctgcc aaaggtaaat taactaaaga tttattcagg aatccccatt tgaatttgta 4501 tgattcaata aaagaaaaca ccaagtaagt tatataaaat aaattgtgta tgagatgttg 4561 tgttttcctt tgtaatttcc actaactaac taactaactt atattcttca tggaatggag 4621 cccagaagaa atgagaggaa gcccttttca cactagatct tatttgaaga aatgtttgtt 4681 agtcagtcag tcagtggttt ctggctctgc cgagggagat gtgttcccca gcaaccattt 4741 ctgcagccca gaatctcaag gcactagagg cggtgtctta attaattggc ttcacaaaga 4801 caaaatgctc tggactggga tttttccttt gctgtgttgg gaatatgtgt ttattaatta 4861 gcacatgcca acaaaataaa tgtcaagagt tatttcataa gtgtaagtaa acttaagaat 4921 taaagagtgc agacttataa ttttca

In some embodiments of the methods of the disclosure, the wild type human FAM13A gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_001252509.1, transcript variant 5):

(SEQ ID NO: 43) 1 maceimplqr llertirsav eqhlfdvnns ggqssedses gtlsassats arqrrrqske 61 qdevrhgrdk glinkentps gfnhlddcil ntqevekvhk ntfgcagers kpkrqksstk 121 lselhdnqdg lvnmeslnst rshertgpdd fewmsderkg nekdgghtqh fesptmkiqe 181 hpslsdtkqq rnqdagdqee sfvsevpqsd ltalcdeknw eepipafssw qrensdsdea 241 hlspqagrli rqlldedsdp mlsprfyayg qsrqylddte vppsppnshs fmrrrssslg 301 syddeqedlt paqltrriqs lkkkirkfed rfeeekkyrp shsdkaanpe vlkwtndlak 361 frrqlkeskl kiseedltpr mrqrsntlpk sfgsqleked ekkqelvdka ikpsveatle 421 siqrklqekr aessrpedik dmtkdqiane kvalqkally yesihgrpvt knerqvmkpl 481 ydryrlvkqi lsrantipii gspsskrrsp llqpiieget asffkeikee eegseddsnv 541 kpdfmvtlkt dfsarcfldq feddadgfis pmddkipskc sqdtglsnlh aasipelleh 601 lqemreekkr irkklrdfed nffrqngrnv qkedrtpmae eyseykhika klrllevlis 661 krdtdsksm

In some embodiments of the methods of the disclosure, the wild type human DSP gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_004415.3, transcript variant 1):

(SEQ ID NO: 44) 1 aagaaaccgg ccaggtgtgg cctaggcgcc cagtgccagc ggggaggaga ctcgctccgc 61 cgccgaccaa caccaacacc cagctccgac gcagctcctc tgcgcccttg ccgccctccg 121 agccacagct ttcctcccgc tcctgccccc ggcccgtcgc cgtctccgcg ctcgcagcgg 181 cctcgggagg gcccaggtag cgagcagcga cctcgcgagc cttccgcact cccgcccggt 241 tccccggccg tccgcctatc cttggccccc tccgctttct ccgcgccggc ccgcctcgct 301 tatgcctcgg cgctgagccg ctctcccgat tgcccgccga catgagctgc aacggaggct 361 cccacccgcg gatcaacact ctgggccgca tgatccgcgc cgagtctggc ccggacctgc 421 gctacgaggt gaccagcggc ggcgggggca ccagcaggat gtactattct cggcgcggcg 481 tgatcaccga ccagaactcg gacggctact gtcaaaccgg cacgatgtcc aggcaccaga 541 accagaacac catccaggag ctgctgcaga actgctccga ctgcttgatg cgagcagagc 601 tcatcgtgca gcctgaattg aagtatggag atggaataca actgactcgg agtcgagaat 661 tggatgagtg ttttgcccag gccaatgacc aaatggaaat cctcgacagc ttgatcagag 721 agatgcggca gatgggccag ccctgtgatg cttaccagaa aaggcttctt cagctccaag 781 agcaaatgcg agccctttat aaagccatca gtgtccctcg agtccgcagg gccagctcca 841 agggtggtgg aggctacact tgtcagagtg gctctggctg ggatgagttc accaaacatg 901 tcaccagtga atgtttgggg tggatgaggc agcaaagggc ggagatggac atggtggcct 961 ggggtgtgga cctggcctca gtggagcagc acattaacag ccaccggggc atccacaact 1021 ccatcggcga ctatcgctgg cagctggaca aaatcaaagc cgacctgcgc gagaaatctg 1081 cgatctacca gttggaggag gagtatgaaa acctgctgaa agcgtccttt gagaggatgg 1141 atcacctgcg acagctgcag aacatcattc aggccacgtc cagggagatc atgtggatca 1201 atgactgcga ggaggaggag ctgctgtacg actggagcga caagaacacc aacatcgctc 1261 agaaacagga ggccttctcc atacgcatga gtcaactgga agttaaagaa aaagagctca 1321 ataagctgaa acaagaaagt gaccaacttg tcctcaatca gcatccagct tcagacaaaa 1381 ttgaggccta tatggacact ctgcagacgc agtggagttg gattcttcag atcaccaagt 1441 gcattgatgt tcatctgaaa gaaaatgctg cctactttca gttttttgaa gaggcgcagt 1501 ctactgaagc atacctgaag gggctccagg actccatcag gaagaagtac ccctgcgaca 1561 agaacatgcc cctgcagcac ctgctggaac agatcaagga gctggagaaa gaacgagaga 1621 aaatccttga atacaagcgt caggtgcaga acttggtaaa caagtctaag aagattgtac 1681 agctgaagcc tcgtaaccca gactacagaa gcaataaacc cattattctc agagctctct 1741 gtgactacaa acaagatcag aaaatcgtgc ataaggggga tgagtgtatc ctgaaggaca 1801 acaacgagcg cagcaagtgg tacgtgacgg gcccgggagg cgttgacatg cttgttccct 1861 ctgtggggct gatcatccct cctccgaacc cactggccgt ggacctctct tgcaagattg 1921 agcagtacta cgaagccatc ttggctctgt ggaaccagct ctacatcaac atgaagagcc 1981 tggtgtcctg gcactactgc atgattgaca tagagaagat cagggccatg acaatcgcca 2041 agctgaaaac aatgcggcag gaagattaca tgaagacgat agccgacctt gagttacatt 2101 accaagagtt catcagaaat agccaaggct cagagatgtt tggagatgat gacaagcgga 2161 aaatacagtc tcagttcacc gatgcccaga agcattacca gaccctggtc attcagctcc 2221 ctggctatcc ccagcaccag acagtgacca caactgaaat cactcatcat ggaacctgcc 2281 aagatgtcaa ccataataaa gtaattgaaa ccaacagaga aaatgacaag caagaaacat 2341 ggatgctgat ggagctgcag aagattcgca ggcagataga gcactgcgag ggcaggatga 2401 ctctcaaaaa cctccctcta gcagaccagg gatcttctca ccacatcaca gtgaaaatta 2461 acgagcttaa gagtgtgcag aatgattcac aagcaattgc tgaggttctc aaccagctta 2521 aagatatgct tgccaacttc agaggttctg aaaagtactg ctatttacag aatgaagtat 2581 ttggactatt tcagaaactg gaaaatatca atggtgttac agatggctac ttaaatagct 2641 tatgcacagt aagggcactg ctccaggcta ttctccaaac agaagacatg ttaaaggttt 2701 atgaagccag gctcactgag gaggaaactg tctgcctgga cctggataaa gtggaagctt 2761 accgctgtgg actgaagaaa ataaaaaatg acttgaactt gaagaagtcg ttgttggcca 2821 ctatgaagac agaactacag aaagcccagc agatccactc tcagacttca cagcagtatc 2881 cactttatga tctggacttg ggcaagttcg gtgaaaaagt cacacagctg acagaccgct 2941 ggcaaaggat agataaacag atcgacttta ggttatggga cctggagaaa caaatcaagc 3001 aattgaggaa ttatcgtgat aactatcagg ctttctgcaa gtggctctat gatgctaaac 3061 gccgccagga ttccttagaa tccatgaaat ttggagattc caacacagtc atgcggtttt 3121 tgaatgagca gaagaacttg cacagtgaaa tatctggcaa acgagacaaa tcagaggaag 3181 tacaaaaaat tgctgaactt tgcgccaatt caattaagga ttatgagctc cagctggcct 3241 catacacctc aggactggaa actctgctga acatacctat caagaggacc atgattcagt 3301 ccccttctgg ggtgattctg caagaggctg cagatgttca tgctcggtac attgaactac 3361 ttacaagatc tggagactat tacaggttct taagtgagat gctgaagagt ttggaagatc 3421 tgaagctgaa aaataccaag atcgaagttt tggaagagga gctcagactg gcccgagatg 3481 ccaactcgga aaactgtaat aagaacaaat tcctggatca gaacctgcag aaataccagg 3541 cagagtgttc ccagttcaaa gcgaagcttg cgagcctgga ggagctgaag agacaggctg 3601 agctggatgg gaagtcggct aagcaaaatc tagacaagtg ctacggccaa ataaaagaac 3661 tcaatgagaa gatcacccga ctgacttatg agattgaaga tgaaaagaga agaagaaaat 3721 ctgtggaaga cagatttgac caacagaaga atgactatga ccaactgcag aaagcaaggc 3781 aatgtgaaaa ggagaacctt ggttggcaga aattagagtc tgagaaagcc atcaaggaga 3841 aggagtacga gattgaaagg ttgagggttc tactgcagga agaaggcacc cggaagagag 3901 aatatgaaaa tgagctggca aaggtaagaa accactataa tgaggagatg agtaatttaa 3961 ggaacaagta tgaaacagag attaacatta cgaagaccac catcaaggag atatccatgc 4021 aaaaagagga tgattccaaa aatcttagaa accagcttga tagactttca agggaaaatc 4081 gagatctgaa ggatgaaatt gtcaggctca atgacagcat cttgcaggcc actgagcagc 4141 gaaggcgagc tgaagaaaac gcccttcagc aaaaggcctg tggctctgag ataatgcaga 4201 agaagcagca tctggagata gaactgaagc aggtcatgca gcagcgctct gaggacaatg 4261 cccggcacaa gcagtccctg gaggaggctg ccaagaccat tcaggacaaa aataaggaga 4321 tcgagagact caaagctgag tttcaggagg aggccaagcg ccgctgggaa tatgaaaatg 4381 aactgagtaa ggtaagaaac aattatgatg aggagatcat tagcttaaaa aatcagtttg 4441 agaccgagat caacatcacc aagaccacca tccaccagct caccatgcag aaggaagagg 4501 ataccagtgg ctaccgggct cagatagaca atctcacccg agaaaacagg agcttatctg 4561 aagaaataaa gaggctgaag aacactctaa cccagaccac agagaatctc aggagggtgg 4621 aagaagacat ccaacagcaa aaggccactg gctctgaggt gtctcagagg aaacagcagc 4681 tggaggttga gctgagacaa gtcactcaga tgcgaacaga ggagagcgta agatataagc 4741 aatctcttga tgatgctgcc aaaaccatcc aggataaaaa caaggagata gaaaggttaa 4801 aacaactgat cgacaaagaa acaaatgacc ggaaatgcct ggaagatgaa aacgcgagat 4861 tacaaagggt ccagtatgac ctgcagaaag caaacagtag tgcgacggag acaataaaca 4921 aactgaaggt tcaggagcaa gaactgacac gcctgaggat cgactatgaa agggtttccc 4981 aggagaggac tgtgaaggac caggatatca cgcggttcca gaactctctg aaagagctgc 5041 agctgcagaa gcagaaggtg gaagaggagc tgaatcggct gaagaggacc gcgtcagaag 5101 actcctgcaa gaggaagaag ctggaggaag agctggaagg catgaggagg tcgctgaagg 5161 agcaagccat caaaatcacc aacctgaccc agcagctgga gcaggcatcc attgttaaga 5221 agaggagtga ggatgacctc cggcagcaga gggacgtgct ggatggccac ctgagggaaa 5281 agcagaggac ccaggaagag ctgaggaggc tctcttctga ggtcgaggcc ctgaggcggc 5341 agttactcca ggaacaggaa agtgtcaaac aagctcactt gaggaatgag catttccaga 5401 aggcgataga agataaaagc agaagcttaa atgaaagcaa aatagaaatt gagaggctgc 5461 agtctctcac agagaacctg accaaggagc acttgatgtt agaagaagaa ctgcggaacc 5521 tgaggctgga gtacgatgac ctgaggagag gacgaagcga agcggacagt gataaaaatg 5581 caaccatctt ggaactaagg agccagctgc agatcagcaa caaccggacc ctggaactgc 5641 aggggctgat taatgattta cagagagaga gggaaaattt gagacaggaa attgagaaat 5701 tccaaaagca ggctttagag gcatctaata ggattcagga atcaaagaat cagtgtactc 5761 aggtggtaca ggaaagagag agccttctgg tgaaaatcaa agtcctggag caagacaagg 5821 caaggctgca gaggctggag gatgagctga atcgtgcaaa atcaactcta gaggcagaaa 5881 ccagggtgaa acagcgcctg gagtgtgaga aacagcaaat tcagaatgac ctgaatcagt 5941 ggaagactca atattcccgc aaggaggagg ctattaggaa gatagaatcg gaaagagaaa 6001 agagtgagag agagaagaac agtcttagga gtgagatcga aagactccaa gcagagatca 6061 agagaattga agagaggtgc aggcgtaagc tggaggattc taccagggag acacagtcac 6121 agttagaaac agaacgctcc cgatatcaga gggagattga taaactcaga cagcgcccat 6181 atgggtccca tcgagagacc cagactgagt gtgagtggac cgttgacacc tccaagctgg 6241 tgtttgatgg gctgaggaag aaggtgacag caatgcagct ctatgagtgt cagctgatcg 6301 acaaaacaac cttggacaaa ctattgaagg ggaagaagtc agtggaagaa gttgcttctg 6361 aaatccagcc attccttcgg ggtgcaggat ctatcgctgg agcatctgct tctcctaagg 6421 aaaaatactc tttggtagag gccaagagaa agaaattaat cagcccagaa tccacagtca 6481 tgcttctgga ggcccaggca gctacaggtg gtataattga tccccatcgg aatgagaagc 6541 tgactgtcga cagtgccata gctcgggacc tcattgactt cgatgaccgt cagcagatat 6601 atgcagcaga aaaagctatc actggttttg atgatccatt ttcaggcaag acagtatctg 6661 tttcagaagc catcaagaaa aatttgattg atagagaaac cggaatgcgc ctgctggaag 6721 cccagattgc ttcagggggt gtagtagacc ctgtgaacag tgtctttttg ccaaaagatg 6781 tcgccttggc ccgggggctg attgatagag atttgtatcg atccctgaat gatccccgag 6841 atagtcagaa aaactttgtg gatccagtca ccaaaaagaa ggtcagttac gtgcagctga 6901 aggaacggtg cagaatcgaa ccacatactg gtctgctctt gctttcagta cagaagagaa 6961 gcatgtcctt ccaaggaatc agacaacctg tgaccgtcac tgagctagta gattctggta 7021 tattgagacc gtccactgtc aatgaactgg aatctggtca gatttcttat gacgaggttg 7081 gtgagagaat taaggacttc ctccagggtt caagctgcat agcaggcata tacaatgaga 7141 ccacaaaaca gaagcttggc atttatgagg ccatgaaaat tggcttagtc cgacctggta 7201 ctgctctgga gttgctggaa gcccaagcag ctactggctt tatagtggat cctgttagca 7261 acttgaggtt accagtggag gaagcctaca agagaggtct ggtgggcatt gagttcaaag 7321 agaagctcct gtctgcagaa cgagctgtca ctgggtataa tgatcctgaa acaggaaaca 7381 tcatctcttt gttccaagcc atgaataagg aactcatcga aaagggccac ggtattcgct 7441 tattagaagc acagatcgca accgggggga tcattgaccc aaaggagagc catcgtttac 7501 cagttgacat agcatataag aggggctatt tcaatgagga actcagtgag attctctcag 7561 atccaagtga tgataccaaa ggattttttg accccaacac tgaagaaaat cttacctatc 7621 tgcaactaaa agaaagatgc attaaggatg aggaaacagg gctctgtctt ctgcctctga 7681 aagaaaagaa gaaacaggtg cagacatcac aaaagaatac cctcaggaag cgtagagtgg 7741 tcatagttga cccagaaacc aataaagaaa tgtctgttca ggaggcctac aagaagggcc 7801 taattgatta tgaaaccttc aaagaactgt gtgagcagga atgtgaatgg gaagaaataa 7861 ccatcacggg atcagatggc tccaccaggg tggtcctggt agatagaaag acaggcagtc 7921 agtatgatat tcaagatgct attgacaagg gccttgttga caggaagttc tttgatcagt 7981 accgatccgg cagcctcagc ctcactcaat ttgctgacat gatctccttg aaaaatggtg 8041 tcggcaccag cagcagcatg ggcagtggtg tcagcgatga tgtttttagc agctcccgac 8101 atgaatcagt aagtaagatt tccaccatat ccagcgtcag gaatttaacc ataaggagca 8161 gctctttttc agacaccctg gaagaatcga gccccattgc agccatcttt gacacagaaa 8221 acctggagaa aatctccatt acagaaggta tagagcgggg catcgttgac agcatcacgg 8281 gtcagaggct tctggaggct caggcctgca caggtggcat catccaccca accacgggcc 8341 agaagctgtc acttcaggac gcagtctccc agggtgtgat tgaccaagac atggccacca 8401 ggctgaagcc tgctcagaaa gccttcatag gcttcgaggg tgtgaaggga aagaagaaga 8461 tgtcagcagc agaggcagtg aaagaaaaat ggctcccgta tgaggctggc cagcgcttcc 8521 tggagttcca gtacctcacg ggaggtcttg ttgacccgga agtgcatggg aggataagca 8581 ccgaagaagc catccggaag gggttcatag atggccgcgc cgcacagagg ctgcaagaca 8641 ccagcagcta tgccaaaatc ctgacctgcc ccaaaaccaa attaaaaata tcctataagg 8701 atgccataaa tcgctccatg gtagaagata tcactgggct gcgccttctg gaagccgcct 8761 ccgtgtcgtc caagggctta cccagccctt acaacatgtc ttcggctccg gggtcccgct 8821 ccggctcccg ctcgggatct cgctccggat ctcgctccgg gtcccgcagt gggtcccgga 8881 gaggaagctt tgacgccaca gggaattctt cctactctta ttcctactca tttagcagta 8941 gttctattgg gcactagtag tcagttggga gtggttgcta taccttgact tcatttatat 9001 gaatttccac tttattaaat aatagaaaag aaaatcccgg tgcttgcagt agagtgatag 9061 gacattctat gcttacagaa aatatagcca tgattgaaat caaatagtaa aggctgttct 9121 ggctttttat cttcttagct catcttaaat aagcagtaca cttggatgca gtgcgtctga 9181 agtgctaatc agttgtaaca atagcacaaa tcgaacttag gatttgtttc ttctcttctg 9241 tgtttcgatt tttgatcaat tctttaattt tggaagccta taatacagtt ttctattctt 9301 ggagataaaa attaaatgga tcactgatat tttagtcatt ctgcttctca tctaaatatt 9361 tccatattct gtattaggag aaaattaccc tcccagcacc agcccccctc tcaaaccccc 9421 aacccaaaac caagcatttt ggaatgagtc tcctttagtt tcagagtgtg gattgtataa 9481 cccatatact cttcgatgta cttgtttggt ttggtattaa tttgactgtg catgacagcg 9541 gcaatctttt ctttggtcaa agttttctgt ttattttgct tgtcatattc gatgtacttt 9601 aaggtgtctt tatgaagttt gctattctgg caataaactt ttagactttt gaagtgtttg 9661 tgttttaatt taatatgttt ataagcatgt ataaacattt agcatatttt tatcataggt 9721 ctaaaaatat ttgtttacta aatacctgtg aagaaatacc attaaaaaac tatttggttc 9781 tgaattctta ctagaaaaaa aa

In some embodiments of the methods of the disclosure, the wild type human DSP gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_004406.2, transcript variant 1):

(SEQ ID NO: 45) 1 mscnggshpr intlgrmira esgpdlryev tsggggtsrm yysrrgvitd qnsdgycqtg 61 tmsrhqnqnt iqellqncsd clmraelivq pelkygdgiq ltrsreldec faqandqmei 121 ldsliremrq mgqpcdayqk rllqlqeqmr alykaisvpr vrrasskggg gytcqsgsgw 181 deftkhvtse clgwmrqqra emdmvawgvd lasveqhins hrgihnsigd yrwqldkika 241 dlreksaiyq leeeyenllk asfermdhlr qlqniiqats reimwindce eeellydwsd 301 kntniaqkqe afsirmsqle vkekelnklk qesdqlvinq hpasdkieay mdtlqtqwsw 361 ilqitkcidv hlkenaayfq ffeeaqstea ylkglqdsir kkypcdknmp lqhlleqike 421 lekerekile ykrqvqnlvn kskkivqlkp rnpdyrsnkp iilralcdyk qdqkivhkgd 481 ecilkdnner skwyvtgpgg vdmlvpsvgl iipppnplav dlsckieqyy eailalwnql 541 yinmkslvsw hycmidieki ramtiaklkt mrqedymkti adlelhyqef irnsqgsemf 601 gdddkrkiqs qftdaqkhyq tiviqlpgyp qhqtvtttei thhgtcqdvn hnkvietnre 661 ndkqetwmlm elqkirrqie hcegrmtlkn lpladqgssh hitvkinelk svqndsgqia 721 evlnqlkdml anfrgsekyc ylqnevfglf qkleningvt dgylnslctv rallqailqt 781 edmlkvyear lteeetvcld ldkveayrcg lkkikndlnl kksllatmkt elqkaqqihs 841 qtsqqyplyd ldlgkfgekv tqltdrwqri dkqidfrlwd lekqikqlrn yrdnyqafck 901 wlydakrrqd slesmkfgds ntvmrflneq knlhseisgk rdkseevqki aelcansikd 961 yelqlasyts gletllnipi krtmiqspsg vilqeaadvh aryielltrs gdyyrflsem 1021 lksledlklk ntkievleee lrlardanse ncnknkfldq nlqkyqaecs qfkaklasle 1081 elkrqaeldg ksakqnldkc ygqikelnek itrltyeied ekrrrksved rfdqqkndyd 1141 qlqkarqcek enlgwqkles ekaikekeye ierlrvllqe egtrkreyen elakvrnhyn 1201 eemsnlrnky eteinitktt ikeismqked dsknlrnqld rlsrenrdlk deivrlndsi 1261 lqateqrrra eenalqqkac gseimqkkqh leielkqvmq qrsednarhk qsleeaakti 1321 qdknkeierl kaefqeeakr rweyenelsk vrnnydeeii slknqfetei nitkttihql 1381 tmqkeedtsg yraqidnltr enrslseeik rlkntltqtt enlrrveedi qqqkatgsev 1441 sqrkqqleve lrqvtqmrte esvrykqsld daaktiqdkn keierlkqli dketndrkcl 1501 edenarlqry qydlqkanss atetinklkv qeqeltrlri dyervsgert vkdqditrfq 1561 nslkelqlqk qkveeelnrl krtasedsck rkkleeeleg mrrslkeqai kitnitqqle 1621 qasivkkrse ddlrqqrdvl dghlrekqrt qeelrrlsse vealrrqllq eqesvkqahl 1681 rnehfqkaie dksrslnesk ieierlqslt enitkehlml eeelrnlrle yddlrrgrse 1741 adsdknatil elrsqlqisn nrtlelqgli ndlgrerenl rqeiekfqkq aleasnriqe 1801 sknqctqvvq eresllvkik vleqdkarlq rledelnrak stleaetrvk qrlecekqqi 1861 qndlnqwktq ysrkeeairk ieserekser eknslrseie rlqaeikrie ercrrkleds 1921 tretqsqlet ersrygreid klrqrpygsh retqtecewt vdtsklvfdg lrkkvtamql 1981 yecqlidktt ldkllkgkks veevaseiqp flrgagsiag asaspkekys lveakrkkli 2041 spestvmlle aqaatggiid phrnekltvd saiardlidf ddrqqiyaae kaitgfddpf 2101 sgktvsvsea ikknlidret gmrlleaqia sggvvdpvns vflpkdvala rglidrdlyr 2161 slndprdsqk nfvdpvtkkk vsyvqlkerc riephtglll lsvqkrsmsf qgirqpvtvt 2221 elvdsgilrp stvnelesgq isydevgeri kdflqgssci agiynettkq klgiyeamki 2281 glvrpgtale lleaqaatgf ivdpvsnlrl pveeaykrgl vgiefkekll saeravtgyn 2341 dpetgniisl fqamnkelie kghgirllea qiatggiidp keshrlpvdi aykrgyfnee 2401 lseilsdpsd dtkgffdpnt eenitylqlk ercikdeetg lcllplkekk kqvqtsqknt 2461 lrkrrvvivd petnkemsvq eaykkglidy etfkelceqe ceweeititg sdgstrvvlv 2521 drktgsqydi qdaidkglvd rkffdqyrsg slsltqfadm islkngvgts ssmgsgvsdd 2581 vfsssrhesv skistissvr nitirsssfs dtleesspia aifdtenlek isitegierg 2641 ivdsitgqrl leaqactggi ihpttgqkls lqdaysqgvi dqdmatrlkp aqkafigfeg 2701 vkgkkkmsaa eavkekwlpy eagqrflefq yltgglvdpe vhgristeea irkgfidgra 2761 aqrlqdtssy akiltcpktk lkisykdain rsmveditgl rlleaasvss kglpspynms 2821 sapgsrsgsr sgsrsgsrsg srsgsrrgsf datgnssysy sysfssssig h

In some embodiments of the methods of the disclosure, the wild type human DSP gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_001008844.2, transcript variant 2):

(SEQ ID NO: 19) 1 aagaaaccgg ccaggtgtgg cctaggcgcc cagtgccagc ggggaggaga ctcgctccgc 61 cgccgaccaa caccaacacc cagctccgac gcagctcctc tgcgcccttg ccgccctccg 121 agccacagct ttcctcccgc tcctgccccc ggcccgtcgc cgtctccgcg ctcgcagcgg 181 cctcgggagg gcccaggtag cgagcagcga cctcgcgagc cttccgcact cccgcccggt 241 tccccggccg tccgcctatc cttggccccc tccgctttct ccgcgccggc ccgcctcgct 301 tatgcctcgg cgctgagccg ctctcccgat tgcccgccga catgagctgc aacggaggct 361 cccacccgcg gatcaacact ctgggccgca tgatccgcgc cgagtctggc ccggacctgc 421 gctacgaggt gaccagcggc ggcgggggca ccagcaggat gtactattct cggcgcggcg 481 tgatcaccga ccagaactcg gacggctact gtcaaaccgg cacgatgtcc aggcaccaga 541 accagaacac catccaggag ctgctgcaga actgctccga ctgcttgatg cgagcagagc 601 tcatcgtgca gcctgaattg aagtatggag atggaataca actgactcgg agtcgagaat 661 tggatgagtg ttttgcccag gccaatgacc aaatggaaat cctcgacagc ttgatcagag 721 agatgcggca gatgggccag ccctgtgatg cttaccagaa aaggcttctt cagctccaag 781 agcaaatgcg agccctttat aaagccatca gtgtccctcg agtccgcagg gccagctcca 841 agggtggtgg aggctacact tgtcagagtg gctctggctg ggatgagttc accaaacatg 901 tcaccagtga atgtttgggg tggatgaggc agcaaagggc ggagatggac atggtggcct 961 ggggtgtgga cctggcctca gtggagcagc acattaacag ccaccggggc atccacaact 1021 ccatcggcga ctatcgctgg cagctggaca aaatcaaagc cgacctgcgc gagaaatctg 1081 cgatctacca gttggaggag gagtatgaaa acctgctgaa agcgtccttt gagaggatgg 1141 atcacctgcg acagctgcag aacatcattc aggccacgtc cagggagatc atgtggatca 1201 atgactgcga ggaggaggag ctgctgtacg actggagcga caagaacacc aacatcgctc 1261 agaaacagga ggccttctcc atacgcatga gtcaactgga agttaaagaa aaagagctca 1321 ataagctgaa acaagaaagt gaccaacttg tcctcaatca gcatccagct tcagacaaaa 1381 ttgaggccta tatggacact ctgcagacgc agtggagttg gattcttcag atcaccaagt 1441 gcattgatgt tcatctgaaa gaaaatgctg cctactttca gttttttgaa gaggcgcagt 1501 ctactgaagc atacctgaag gggctccagg actccatcag gaagaagtac ccctgcgaca 1561 agaacatgcc cctgcagcac ctgctggaac agatcaagga gctggagaaa gaacgagaga 1621 aaatccttga atacaagcgt caggtgcaga acttggtaaa caagtctaag aagattgtac 1681 agctgaagcc tcgtaaccca gactacagaa gcaataaacc cattattctc agagctctct 1741 gtgactacaa acaagatcag aaaatcgtgc ataaggggga tgagtgtatc ctgaaggaca 1801 acaacgagcg cagcaagtgg tacgtgacgg gcccgggagg cgttgacatg cttgttccct 1861 ctgtggggct gatcatccct cctccgaacc cactggccgt ggacctctct tgcaagattg 1921 agcagtacta cgaagccatc ttggctctgt ggaaccagct ctacatcaac atgaagagcc 1981 tggtgtcctg gcactactgc atgattgaca tagagaagat cagggccatg acaatcgcca 2041 agctgaaaac aatgcggcag gaagattaca tgaagacgat agccgacctt gagttacatt 2101 accaagagtt catcagaaat agccaaggct cagagatgtt tggagatgat gacaagcgga 2161 aaatacagtc tcagttcacc gatgcccaga agcattacca gaccctggtc attcagctcc 2221 ctggctatcc ccagcaccag acagtgacca caactgaaat cactcatcat ggaacctgcc 2281 aagatgtcaa ccataataaa gtaattgaaa ccaacagaga aaatgacaag caagaaacat 2341 ggatgctgat ggagctgcag aagattcgca ggcagataga gcactgcgag ggcaggatga 2401 ctctcaaaaa cctccctcta gcagaccagg gatcttctca ccacatcaca gtgaaaatta 2461 acgagcttaa gagtgtgcag aatgattcac aagcaattgc tgaggttctc aaccagctta 2521 aagatatgct tgccaacttc agaggttctg aaaagtactg ctatttacag aatgaagtat 2581 ttggactatt tcagaaactg gaaaatatca atggtgttac agatggctac ttaaatagct 2641 tatgcacagt aagggcactg ctccaggcta ttctccaaac agaagacatg ttaaaggttt 2701 atgaagccag gctcactgag gaggaaactg tctgcctgga cctggataaa gtggaagctt 2761 accgctgtgg actgaagaaa ataaaaaatg acttgaactt gaagaagtcg ttgttggcca 2821 ctatgaagac agaactacag aaagcccagc agatccactc tcagacttca cagcagtatc 2881 cactttatga tctggacttg ggcaagttcg gtgaaaaagt cacacagctg acagaccgct 2941 ggcaaaggat agataaacag atcgacttta ggttatggga cctggagaaa caaatcaagc 3001 aattgaggaa ttatcgtgat aactatcagg ctttctgcaa gtggctctat gatgctaaac 3061 gccgccagga ttccttagaa tccatgaaat ttggagattc caacacagtc atgcggtttt 3121 tgaatgagca gaagaacttg cacagtgaaa tatctggcaa acgagacaaa tcagaggaag 3181 tacaaaaaat tgctgaactt tgcgccaatt caattaagga ttatgagctc cagctggcct 3241 catacacctc aggactggaa actctgctga acatacctat caagaggacc atgattcagt 3301 ccccttctgg ggtgattctg caagaggctg cagatgttca tgctcggtac attgaactac 3361 ttacaagatc tggagactat tacaggttct taagtgagat gctgaagagt ttggaagatc 3421 tgaagctgaa aaataccaag atcgaagttt tggaagagga gctcagactg gcccgagatg 3481 ccaactcgga aaactgtaat aagaacaaat tcctggatca gaacctgcag aaataccagg 3541 cagagtgttc ccagttcaaa gcgaagcttg cgagcctgga ggagctgaag agacaggctg 3601 agctggatgg gaagtcggct aagcaaaatc tagacaagtg ctacggccaa ataaaagaac 3661 tcaatgagaa gatcacccga ctgacttatg agattgaaga tgaaaagaga agaagaaaat 3721 ctgtggaaga cagatttgac caacagaaga atgactatga ccaactgcag aaagcaaggc 3781 aatgtgaaaa ggagaacctt ggttggcaga aattagagtc tgagaaagcc atcaaggaga 3841 aggagtacga gattgaaagg ttgagggttc tactgcagga agaaggcacc cggaagagag 3901 aatatgaaaa tgagctggca aaggcatcta ataggattca ggaatcaaag aatcagtgta 3961 ctcaggtggt acaggaaaga gagagccttc tggtgaaaat caaagtcctg gagcaagaca 4021 aggcaaggct gcagaggctg gaggatgagc tgaatcgtgc aaaatcaact ctagaggcag 4081 aaaccagggt gaaacagcgc ctggagtgtg agaaacagca aattcagaat gacctgaatc 4141 agtggaagac tcaatattcc cgcaaggagg aggctattag gaagatagaa tcggaaagag 4201 aaaagagtga gagagagaag aacagtctta ggagtgagat cgaaagactc caagcagaga 4261 tcaagagaat tgaagagagg tgcaggcgta agctggagga ttctaccagg gagacacagt 4321 cacagttaga aacagaacgc tcccgatatc agagggagat tgataaactc agacagcgcc 4381 catatgggtc ccatcgagag acccagactg agtgtgagtg gaccgttgac acctccaagc 4441 tggtgtttga tgggctgagg aagaaggtga cagcaatgca gctctatgag tgtcagctga 4501 tcgacaaaac aaccttggac aaactattga aggggaagaa gtcagtggaa gaagttgctt 4561 ctgaaatcca gccattcctt cggggtgcag gatctatcgc tggagcatct gcttctccta 4621 aggaaaaata ctctttggta gaggccaaga gaaagaaatt aatcagccca gaatccacag 4681 tcatgcttct ggaggcccag gcagctacag gtggtataat tgatccccat cggaatgaga 4741 agctgactgt cgacagtgcc atagctcggg acctcattga cttcgatgac cgtcagcaga 4801 tatatgcagc agaaaaagct atcactggtt ttgatgatcc attttcaggc aagacagtat 4861 ctgtttcaga agccatcaag aaaaatttga ttgatagaga aaccggaatg cgcctgctgg 4921 aagcccagat tgcttcaggg ggtgtagtag accctgtgaa cagtgtcttt ttgccaaaag 4981 atgtcgcctt ggcccggggg ctgattgata gagatttgta tcgatccctg aatgatcccc 5041 gagatagtca gaaaaacttt gtggatccag tcaccaaaaa gaaggtcagt tacgtgcagc 5101 tgaaggaacg gtgcagaatc gaaccacata ctggtctgct cttgctttca gtacagaaga 5161 gaagcatgtc cttccaagga atcagacaac ctgtgaccgt cactgagcta gtagattctg 5221 gtatattgag accgtccact gtcaatgaac tggaatctgg tcagatttct tatgacgagg 5281 ttggtgagag aattaaggac ttcctccagg gttcaagctg catagcaggc atatacaatg 5341 agaccacaaa acagaagctt ggcatttatg aggccatgaa aattggctta gtccgacctg 5401 gtactgctct ggagttgctg gaagcccaag cagctactgg ctttatagtg gatcctgtta 5461 gcaacttgag gttaccagtg gaggaagcct acaagagagg tctggtgggc attgagttca 5521 aagagaagct cctgtctgca gaacgagctg tcactgggta taatgatcct gaaacaggaa 5581 acatcatctc tttgttccaa gccatgaata aggaactcat cgaaaagggc cacggtattc 5641 gcttattaga agcacagatc gcaaccgggg ggatcattga cccaaaggag agccatcgtt 5701 taccagttga catagcatat aagaggggct atttcaatga ggaactcagt gagattctct 5761 cagatccaag tgatgatacc aaaggatttt ttgaccccaa cactgaagaa aatcttacct 5821 atctgcaact aaaagaaaga tgcattaagg atgaggaaac agggctctgt cttctgcctc 5881 tgaaagaaaa gaagaaacag gtgcagacat cacaaaagaa taccctcagg aagcgtagag 5941 tggtcatagt tgacccagaa accaataaag aaatgtctgt tcaggaggcc tacaagaagg 6001 gcctaattga ttatgaaacc ttcaaagaac tgtgtgagca ggaatgtgaa tgggaagaaa 6061 taaccatcac gggatcagat ggctccacca gggtggtcct ggtagataga aagacaggca 6121 gtcagtatga tattcaagat gctattgaca agggccttgt tgacaggaag ttctttgatc 6181 agtaccgatc cggcagcctc agcctcactc aatttgctga catgatctcc ttgaaaaatg 6241 gtgtcggcac cagcagcagc atgggcagtg gtgtcagcga tgatgttttt agcagctccc 6301 gacatgaatc agtaagtaag atttccacca tatccagcgt caggaattta accataagga 6361 gcagctcttt ttcagacacc ctggaagaat cgagccccat tgcagccatc tttgacacag 6421 aaaacctgga gaaaatctcc attacagaag gtatagagcg gggcatcgtt gacagcatca 6481 cgggtcagag gcttctggag gctcaggcct gcacaggtgg catcatccac ccaaccacgg 6541 gccagaagct gtcacttcag gacgcagtct cccagggtgt gattgaccaa gacatggcca 6601 ccaggctgaa gcctgctcag aaagccttca taggcttcga gggtgtgaag ggaaagaaga 6661 agatgtcagc agcagaggca gtgaaagaaa aatggctccc gtatgaggct ggccagcgct 6721 tcctggagtt ccagtacctc acgggaggtc ttgttgaccc ggaagtgcat gggaggataa 6781 gcaccgaaga agccatccgg aaggggttca tagatggccg cgccgcacag aggctgcaag 6841 acaccagcag ctatgccaaa atcctgacct gccccaaaac caaattaaaa atatcctata 6901 aggatgccat aaatcgctcc atggtagaag atatcactgg gctgcgcctt ctggaagccg 6961 cctccgtgtc gtccaagggc ttacccagcc cttacaacat gtcttcggct ccggggtccc 7021 gctccggctc ccgctcggga tctcgctccg gatctcgctc cgggtcccgc agtgggtccc 7081 ggagaggaag ctttgacgcc acagggaatt cttcctactc ttattcctac tcatttagca 7141 gtagttctat tgggcactag tagtcagttg ggagtggttg ctataccttg acttcattta 7201 tatgaatttc cactttatta aataatagaa aagaaaatcc cggtgcttgc agtagagtga 7261 taggacattc tatgcttaca gaaaatatag ccatgattga aatcaaatag taaaggctgt 7321 tctggctttt tatcttctta gctcatctta aataagcagt acacttggat gcagtgcgtc 7381 tgaagtgcta atcagttgta acaatagcac aaatcgaact taggatttgt ttcttctctt 7441 ctgtgtttcg atttttgatc aattctttaa ttttggaagc ctataataca gttttctatt 7501 cttggagata aaaattaaat ggatcactga tattttagtc attctgcttc tcatctaaat 7561 atttccatat tctgtattag gagaaaatta ccctcccagc accagccccc ctctcaaacc 7621 cccaacccaa aaccaagcat tttggaatga gtctccttta gtttcagagt gtggattgta 7681 taacccatat actcttcgat gtacttgttt ggtttggtat taatttgact gtgcatgaca 7741 gcggcaatct tttctttggt caaagttttc tgtttatttt gcttgtcata ttcgatgtac 7801 tttaaggtgt ctttatgaag tttgctattc tggcaataaa cttttagact tttgaagtgt 7861 ttgtgtttta atttaatatg tttataagca tgtataaaca tttagcatat ttttatcata 7921 ggtctaaaaa tatttgttta ctaaatacct gtgaagaaat accattaaaa aactatttgg 7981 ttctgaattc ttactagaaa aaaaa

In some embodiments of the methods of the disclosure, the wild type human DSP gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_001008844.1, transcript variant 2):

(SEQ ID NO: 20) 1 mscnggshpr intlgrmira esgpdlryev tsggggtsrm yysrrgvitd qnsdgycqtg 61 tmsrhqnqnt iqellqncsd clmraelivq pelkygdgiq ltrsreldec faqandqmei 121 ldsliremrq mgqpcdayqk rllqlqeqmr alykaisvpr vrrasskggg gytcqsgsgw 181 deftkhvtse clgwmrqqra emdmvawgvd lasveqhins hrgihnsigd yrwqldkika 241 dlreksaiyq leeeyenllk asfermdhlr qlqniiqats reimwindce eeellydwsd 301 kntniaqkqe afsirmsqle vkekelnklk qesdqlvinq hpasdkieay mdtlqtqwsw 361 ilqitkcidv hlkenaayfq ffeeaqstea ylkglqdsir kkypcdknmp lqhlleqike 421 lekerekile ykrqvqnlvn kskkivqlkp rnpdyrsnkp iilralcdyk qdqkivhkgd 481 ecilkdnner skwyvtgpgg vdmlvpsvgl iipppnplav dlsckieqyy eailalwnql 541 yinmkslvsw hycmidieki ramtiaklkt mrqedymkti adlelhyqef irnsqgsemf 601 gdddkrkiqs qftdaqkhyq tiviqlpgyp qhqtvtttei thhgtcqdvn hnkvietnre 661 ndkqetwmlm elqkirrqie hcegrmtlkn lpladqgssh hitvkinelk svqndsgaia 721 evinqlkdml anfrgsekyc ylqnevfglf qkleningvt dgylnslctv rallqailqt 781 edmlkvyear lteeetvcld ldkveayrcg lkkikndlnl kksllatmkt elqkaqqihs 841 qtsqqyplyd ldlgkfgekv tqltdrwqri dkqidfrlwd lekqikqlrn yrdnyqafck 901 wlydakrrqd slesmkfgds ntvmrflneq knlhseisgk rdkseevqki aelcansikd 961 yelqlasyts gletllnipi krtmiqspsg vilqeaadvh aryielltrs gdyyrflsem 1021 lksledlklk ntkievleee lrlardanse ncnknkfldq nlqkyqaecs qfkaklasle 1081 elkrqaeldg ksakqnldkc ygqikelnek itrltyeied ekrrrksved rfdqqkndyd 1141 qlqkarqcek enlgwqkles ekaikekeye ierlrvllqe egtrkreyen elakasnriq 1201 esknqctqvv qeresllvki kvleqdkarl qrledelnra kstleaetrv kqrlecekqq 1261 iqndlnqwkt qysrkeeair kieserekse reknslrsei erlqaeikri eercrrkled 1321 stretqsqle tersrygrei dklrqrpygs hretqtecew tvdtsklvfd glrkkvtamq 1381 lyecqlidkt tldkllkgkk sveevaseiq pflrgagsia gasaspkeky slveakrkkl 1441 ispestvmll eaqaatggii dphrnekltv dsaiardlid fddrqqiyaa ekaitgfddp 1501 fsgktvsvse aikknlidre tgmrlleaqi asggvvdpvn svflpkdval arglidrdly 1561 rslndprdsq knfvdpvtkk kvsyvqlker criephtgll llsvqkrsms fqgirqpvtv 1621 telvdsgilr pstvnelesg qisydevger ikdflqgssc iagiynettk qklgiyeamk 1681 iglvrpgtal elleaqaatg fivdpvsnlr lpveeaykrg lvgiefkekl lsaeravtgy 1741 ndpetgniis lfqamnkeli ekghgirlle aqiatggiid pkeshrlpvd iaykrgyfne 1801 elseilsdps ddtkgffdpn teenitylql kercikdeet glcllplkek kkqvqtsqkn 1861 tlrkrrvviv dpetnkemsv qeaykkglid yetfkelceq eceweeitit gsdgstrvvl 1921 vdrktgsqyd iqdaidkglv drkffdqyrs gslsltqfad mislkngvgt sssmgsgvsd 1981 dvfsssrhes vskistissv rnltirsssf sdtleesspi aaifdtenle kisitegier 2041 givdsitgqr lleaqactgg iihpttgqkl slqdavsqgv idqdmatrlk paqkafigfe 2101 gvkgkkkmsa aeavkekwlp yeagqrflef qyltgglvdp evhgristee airkgfidgr 2161 aaqrlqdtss yakiltcpkt klkisykdai nrsmveditg lrlleaasvs skglpspynm 2221 ssapgsrsgs rsgsrsgsrs gsrsgsrrgs fdatgnssys ysysfssssi gh

In some embodiments of the methods of the disclosure, the wild type human DSP gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_001319034.1, transcript variant 3):

(SEQ ID NO: 46) 1 aagaaaccgg ccaggtgtgg cctaggcgcc cagtgccagc ggggaggaga ctcgctccgc 61 cgccgaccaa caccaacacc cagctccgac gcagctcctc tgcgcccttg ccgccctccg 121 agccacagct ttcctcccgc tcctgccccc ggcccgtcgc cgtctccgcg ctcgcagcgg 181 cctcgggagg gcccaggtag cgagcagcga cctcgcgagc cttccgcact cccgcccggt 241 tccccggccg tccgcctatc cttggccccc tccgctttct ccgcgccggc ccgcctcgct 301 tatgcctcgg cgctgagccg ctctcccgat tgcccgccga catgagctgc aacggaggct 361 cccacccgcg gatcaacact ctgggccgca tgatccgcgc cgagtctggc ccggacctgc 421 gctacgaggt gaccagcggc ggcgggggca ccagcaggat gtactattct cggcgcggcg 481 tgatcaccga ccagaactcg gacggctact gtcaaaccgg cacgatgtcc aggcaccaga 541 accagaacac catccaggag ctgctgcaga actgctccga ctgcttgatg cgagcagagc 601 tcatcgtgca gcctgaattg aagtatggag atggaataca actgactcgg agtcgagaat 661 tggatgagtg ttttgcccag gccaatgacc aaatggaaat cctcgacagc ttgatcagag 721 agatgcggca gatgggccag ccctgtgatg cttaccagaa aaggcttctt cagctccaag 781 agcaaatgcg agccctttat aaagccatca gtgtccctcg agtccgcagg gccagctcca 841 agggtggtgg aggctacact tgtcagagtg gctctggctg ggatgagttc accaaacatg 901 tcaccagtga atgtttgggg tggatgaggc agcaaagggc ggagatggac atggtggcct 961 ggggtgtgga cctggcctca gtggagcagc acattaacag ccaccggggc atccacaact 1021 ccatcggcga ctatcgctgg cagctggaca aaatcaaagc cgacctgcgc gagaaatctg 1081 cgatctacca gttggaggag gagtatgaaa acctgctgaa agcgtccttt gagaggatgg 1141 atcacctgcg acagctgcag aacatcattc aggccacgtc cagggagatc atgtggatca 1201 atgactgcga ggaggaggag ctgctgtacg actggagcga caagaacacc aacatcgctc 1261 agaaacagga ggccttctcc atacgcatga gtcaactgga agttaaagaa aaagagctca 1321 ataagctgaa acaagaaagt gaccaacttg tcctcaatca gcatccagct tcagacaaaa 1381 ttgaggccta tatggacact ctgcagacgc agtggagttg gattcttcag atcaccaagt 1441 gcattgatgt tcatctgaaa gaaaatgctg cctactttca gttttttgaa gaggcgcagt 1501 ctactgaagc atacctgaag gggctccagg actccatcag gaagaagtac ccctgcgaca 1561 agaacatgcc cctgcagcac ctgctggaac agatcaagga gctggagaaa gaacgagaga 1621 aaatccttga atacaagcgt caggtgcaga acttggtaaa caagtctaag aagattgtac 1681 agctgaagcc tcgtaaccca gactacagaa gcaataaacc cattattctc agagctctct 1741 gtgactacaa acaagatcag aaaatcgtgc ataaggggga tgagtgtatc ctgaaggaca 1801 acaacgagcg cagcaagtgg tacgtgacgg gcccgggagg cgttgacatg cttgttccct 1861 ctgtggggct gatcatccct cctccgaacc cactggccgt ggacctctct tgcaagattg 1921 agcagtacta cgaagccatc ttggctctgt ggaaccagct ctacatcaac atgaagagcc 1981 tggtgtcctg gcactactgc atgattgaca tagagaagat cagggccatg acaatcgcca 2041 agctgaaaac aatgcggcag gaagattaca tgaagacgat agccgacctt gagttacatt 2101 accaagagtt catcagaaat agccaaggct cagagatgtt tggagatgat gacaagcgga 2161 aaatacagtc tcagttcacc gatgcccaga agcattacca gaccctggtc attcagctcc 2221 ctggctatcc ccagcaccag acagtgacca caactgaaat cactcatcat ggaacctgcc 2281 aagatgtcaa ccataataaa gtaattgaaa ccaacagaga aaatgacaag caagaaacat 2341 ggatgctgat ggagctgcag aagattcgca ggcagataga gcactgcgag ggcaggatga 2401 ctctcaaaaa cctccctcta gcagaccagg gatcttctca ccacatcaca gtgaaaatta 2461 acgagcttaa gagtgtgcag aatgattcac aagcaattgc tgaggttctc aaccagctta 2521 aagatatgct tgccaacttc agaggttctg aaaagtactg ctatttacag aatgaagtat 2581 ttggactatt tcagaaactg gaaaatatca atggtgttac agatggctac ttaaatagct 2641 tatgcacagt aagggcactg ctccaggcta ttctccaaac agaagacatg ttaaaggttt 2701 atgaagccag gctcactgag gaggaaactg tctgcctgga cctggataaa gtggaagctt 2761 accgctgtgg actgaagaaa ataaaaaatg acttgaactt gaagaagtcg ttgttggcca 2821 ctatgaagac agaactacag aaagcccagc agatccactc tcagacttca cagcagtatc 2881 cactttatga tctggacttg ggcaagttcg gtgaaaaagt cacacagctg acagaccgct 2941 ggcaaaggat agataaacag atcgacttta ggttatggga cctggagaaa caaatcaagc 3001 aattgaggaa ttatcgtgat aactatcagg ctttctgcaa gtggctctat gatgctaaac 3061 gccgccagga ttccttagaa tccatgaaat ttggagattc caacacagtc atgcggtttt 3121 tgaatgagca gaagaacttg cacagtgaaa tatctggcaa acgagacaaa tcagaggaag 3181 tacaaaaaat tgctgaactt tgcgccaatt caattaagga ttatgagctc cagctggcct 3241 catacacctc aggactggaa actctgctga acatacctat caagaggacc atgattcagt 3301 ccccttctgg ggtgattctg caagaggctg cagatgttca tgctcggtac attgaactac 3361 ttacaagatc tggagactat tacaggttct taagtgagat gctgaagagt ttggaagatc 3421 tgaagctgaa aaataccaag atcgaagttt tggaagagga gctcagactg gcccgagatg 3481 ccaactcgga aaactgtaat aagaacaaat tcctggatca gaacctgcag aaataccagg 3541 cagagtgttc ccagttcaaa gcgaagcttg cgagcctgga ggagctgaag agacaggctg 3601 agctggatgg gaagtcggct aagcaaaatc tagacaagtg ctacggccaa ataaaagaac 3661 tcaatgagaa gatcacccga ctgacttatg agattgaaga tgaaaagaga agaagaaaat 3721 ctgtggaaga cagatttgac caacagaaga atgactatga ccaactgcag aaagcaaggc 3781 aatgtgaaaa ggagaacctt ggttggcaga aattagagtc tgagaaagcc atcaaggaga 3841 aggagtacga gattgaaagg ttgagggttc tactgcagga agaaggcacc cggaagagag 3901 aatatgaaaa tgagctggca aaggtaagaa accactataa tgaggagatg agtaatttaa 3961 ggaacaagta tgaaacagag attaacatta cgaagaccac catcaaggag atatccatgc 4021 aaaaagagga tgattccaaa aatcttagaa accagcttga tagactttca agggaaaatc 4081 gagatctgaa ggatgaaatt gtcaggctca atgacagcat cttgcaggcc actgagcagc 4141 gaaggcgagc tgaagaaaac gcccttcagc aaaaggcctg tggctctgag ataatgcaga 4201 agaagcagca tctggagata gaactgaagc aggtcatgca gcagcgctct gaggacaatg 4261 cccggcacaa gcagtccctg gaggaggctg ccaagaccat tcaggacaaa aataaggaga 4321 tcgagagact caaagctgag tttcaggagg aggccaagcg ccgctgggaa tatgaaaatg 4381 aactgagtaa ggcatctaat aggattcagg aatcaaagaa tcagtgtact caggtggtac 4441 aggaaagaga gagccttctg gtgaaaatca aagtcctgga gcaagacaag gcaaggctgc 4501 agaggctgga ggatgagctg aatcgtgcaa aatcaactct agaggcagaa accagggtga 4561 aacagcgcct ggagtgtgag aaacagcaaa ttcagaatga cctgaatcag tggaagactc 4621 aatattcccg caaggaggag gctattagga agatagaatc ggaaagagaa aagagtgaga 4681 gagagaagaa cagtcttagg agtgagatcg aaagactcca agcagagatc aagagaattg 4741 aagagaggtg caggcgtaag ctggaggatt ctaccaggga gacacagtca cagttagaaa 4801 cagaacgctc ccgatatcag agggagattg ataaactcag acagcgccca tatgggtccc 4861 atcgagagac ccagactgag tgtgagtgga ccgttgacac ctccaagctg gtgtttgatg 4921 ggctgaggaa gaaggtgaca gcaatgcagc tctatgagtg tcagctgatc gacaaaacaa 4981 ccttggacaa actattgaag gggaagaagt cagtggaaga agttgcttct gaaatccagc 5041 cattccttcg gggtgcagga tctatcgctg gagcatctgc ttctcctaag gaaaaatact 5101 ctttggtaga ggccaagaga aagaaattaa tcagcccaga atccacagtc atgcttctgg 5161 aggcccaggc agctacaggt ggtataattg atccccatcg gaatgagaag ctgactgtcg 5221 acagtgccat agctcgggac ctcattgact tcgatgaccg tcagcagata tatgcagcag 5281 aaaaagctat cactggtttt gatgatccat tttcaggcaa gacagtatct gtttcagaag 5341 ccatcaagaa aaatttgatt gatagagaaa ccggaatgcg cctgctggaa gcccagattg 5401 cttcaggggg tgtagtagac cctgtgaaca gtgtcttttt gccaaaagat gtcgccttgg 5461 cccgggggct gattgataga gatttgtatc gatccctgaa tgatccccga gatagtcaga 5521 aaaactttgt ggatccagtc accaaaaaga aggtcagtta cgtgcagctg aaggaacggt 5581 gcagaatcga accacatact ggtctgctct tgctttcagt acagaagaga agcatgtcct 5641 tccaaggaat cagacaacct gtgaccgtca ctgagctagt agattctggt atattgagac 5701 cgtccactgt caatgaactg gaatctggtc agatttctta tgacgaggtt ggtgagagaa 5761 ttaaggactt cctccagggt tcaagctgca tagcaggcat atacaatgag accacaaaac 5821 agaagcttgg catttatgag gccatgaaaa ttggcttagt ccgacctggt actgctctgg 5881 agttgctgga agcccaagca gctactggct ttatagtgga tcctgttagc aacttgaggt 5941 taccagtgga ggaagcctac aagagaggtc tggtgggcat tgagttcaaa gagaagctcc 6001 tgtctgcaga acgagctgtc actgggtata atgatcctga aacaggaaac atcatctctt 6061 tgttccaagc catgaataag gaactcatcg aaaagggcca cggtattcgc ttattagaag 6121 cacagatcgc aaccgggggg atcattgacc caaaggagag ccatcgttta ccagttgaca 6181 tagcatataa gaggggctat ttcaatgagg aactcagtga gattctctca gatccaagtg 6241 atgataccaa aggatttttt gaccccaaca ctgaagaaaa tcttacctat ctgcaactaa 6301 aagaaagatg cattaaggat gaggaaacag ggctctgtct tctgcctctg aaagaaaaga 6361 agaaacaggt gcagacatca caaaagaata ccctcaggaa gcgtagagtg gtcatagttg 6421 acccagaaac caataaagaa atgtctgttc aggaggccta caagaagggc ctaattgatt 6481 atgaaacctt caaagaactg tgtgagcagg aatgtgaatg ggaagaaata accatcacgg 6541 gatcagatgg ctccaccagg gtggtcctgg tagatagaaa gacaggcagt cagtatgata 6601 ttcaagatgc tattgacaag ggccttgttg acaggaagtt ctttgatcag taccgatccg 6661 gcagcctcag cctcactcaa tttgctgaca tgatctcctt gaaaaatggt gtcggcacca 6721 gcagcagcat gggcagtggt gtcagcgatg atgtttttag cagctcccga catgaatcag 6781 taagtaagat ttccaccata tccagcgtca ggaatttaac cataaggagc agctcttttt 6841 cagacaccct ggaagaatcg agccccattg cagccatctt tgacacagaa aacctggaga 6901 aaatctccat tacagaaggt atagagcggg gcatcgttga cagcatcacg ggtcagaggc 6961 ttctggaggc tcaggcctgc acaggtggca tcatccaccc aaccacgggc cagaagctgt 7021 cacttcagga cgcagtctcc cagggtgtga ttgaccaaga catggccacc aggctgaagc 7081 ctgctcagaa agccttcata ggcttcgagg gtgtgaaggg aaagaagaag atgtcagcag 7141 cagaggcagt gaaagaaaaa tggctcccgt atgaggctgg ccagcgcttc ctggagttcc 7201 agtacctcac gggaggtctt gttgacccgg aagtgcatgg gaggataagc accgaagaag 7261 ccatccggaa ggggttcata gatggccgcg ccgcacagag gctgcaagac accagcagct 7321 atgccaaaat cctgacctgc cccaaaacca aattaaaaat atcctataag gatgccataa 7381 atcgctccat ggtagaagat atcactgggc tgcgccttct ggaagccgcc tccgtgtcgt 7441 ccaagggctt acccagccct tacaacatgt cttcggctcc ggggtcccgc tccggctccc 7501 gctcgggatc tcgctccgga tctcgctccg ggtcccgcag tgggtcccgg agaggaagct 7561 ttgacgccac agggaattct tcctactctt attcctactc atttagcagt agttctattg 7621 ggcactagta gtcagttggg agtggttgct ataccttgac ttcatttata tgaatttcca 7681 ctttattaaa taatagaaaa gaaaatcccg gtgcttgcag tagagtgata ggacattcta 7741 tgcttacaga aaatatagcc atgattgaaa tcaaatagta aaggctgttc tggcttttta 7801 tcttcttagc tcatcttaaa taagcagtac acttggatgc agtgcgtctg aagtgctaat 7861 cagttgtaac aatagcacaa atcgaactta ggatttgttt cttctcttct gtgtttcgat 7921 ttttgatcaa ttctttaatt ttggaagcct ataatacagt tttctattct tggagataaa 7981 aattaaatgg atcactgata ttttagtcat tctgcttctc atctaaatat ttccatattc 8041 tgtattagga gaaaattacc ctcccagcac cagcccccct ctcaaacccc caacccaaaa 8101 ccaagcattt tggaatgagt ctcctttagt ttcagagtgt ggattgtata acccatatac 8161 tcttcgatgt acttgtttgg tttggtatta atttgactgt gcatgacagc ggcaatcttt 8221 tctttggtca aagttttctg tttattttgc ttgtcatatt cgatgtactt taaggtgtct 8281 ttatgaagtt tgctattctg gcaataaact tttagacttt tgaagtgttt gtgttttaat 8341 ttaatatgtt tataagcatg tataaacatt tagcatattt ttatcatagg tctaaaaata 8401 tttgtttact aaatacctgt gaagaaatac cattaaaaaa ctatttggtt ctgaattctt 8461 actagaaaaa aaa

In some embodiments of the methods of the disclosure, the wild type human DSP gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_001305963.1, transcript variant 3):

(SEQ ID NO: 47)    1 mscnggshpr intlgrmira esgpdlryev tsggggtsrm yysrrgvitd qnsdgycqtg   61 tmsrhqnqnt iqellqncsd clmraelivq pelkygdgiq ltrsreldec faqandqmei  121 ldsliremrq mgqpcdayqk rllqlqeqmr alykaisvpr vrrasskggg gytcqsgsgw  181 deftkhvtse clgwmrqqra emdmvawgvd lasveqhins hrgihnsigd yrwqldkika  241 dlreksaiyq leeeyenllk asfermdhlr qlqniiqats reimwindce eeellydwsd  301 kntniaqkqe afsirmsqle vkekelnklk qesdqlvlnq hpasdkieay mdtlqtqwsw  361 ilqitkcidv hlkenaayfq ffeeaqstea ylkglqdsir kkypcdknmp lqhlleqike  421 lekerekile ykrqvqnlvn kskkivqlkp rnpdyrsnkp iilralcdyk qdqkivhkgd  481 ecilkdnner skwyvtgpgg vdmlvpsvgl iipppnplav dlsckieqyy eailalwnql  541 yinmkslvsw hycmidieki ramtiaklkt mrqedymkti adlelhyqef irnsqgsemf  601 gdddkrkiqs qftdaqkhyq tlviqlpgyp qhqtvtttei thhgtcqdvn hnkvietnre  661 ndkqetwmlm elqkirrqie hcegrmtlkn lpladqgssh hitvkinelk svqndsqaia  721 evlnqlkdml anfrgsekyc ylqnevfglf qkleningvt dgylnslctv rallqailqt  781 edmlkvyear lteeetvcld ldkveayrcg lkkikndlnl kksllatmkt elqkaqqihs  841 qtsqqyplyd ldlgkfgekv tqltdrwqri dkqidfrlwd lekqikqlrn yrdnyqafck  901 wlydakrrqd slesmkfgds ntvmrflneq knlhseisgk rdkseevqki aelcansikd  961 yelqlasyts gletllnipi krtmiqspsg vilqeaadvh aryielltrs gdyyrflsem 1021 lksledlklk ntkievleee lrlardanse ncnknkfldq nlqkyqaecs qfkaklasle 1081 elkrqaeldg ksakqnldkc ygqikelnek itrltyeied ekrrrksved rfdqqkndyd 1141 qlqkarqcek enlgwqkles ekaikekeye ierlrvllqe egtrkreyen elakvrnhyn 1201 eemsnlrnky eteinitktt ikeismqked dsknlrnqld rlsrenrdlk deivrlndsi 1261 lqateqrrra eenalqqkac gseimqkkqh leielkqvmq qrsednarhk qsleeaakti 1321 qdknkeierl kaefqeeakr rweyenelsk asnriqeskn qctqvvqere sllvkikvle 1381 qdkarlqrle delnrakstl eaetrvkqrl ecekqqiqnd lnqwktqysr keeairkies 1441 erekserekn slrseierlq aeikrieerc rrkledstre tqsqleters ryqreidklr 1501 qrpygshret qtecewtvdt sklvfdglrk kvtamqlyec qlidkttldk llkgkksvee 1561 vaseiqpflr gagsiagasa spkekyslve akrkklispe stvmlleaqa atggiidphr 1621 nekltvdsai ardlidfddr qqiyaaekai tgfddpfsgk tvsyseaikk nlidretgmr 1681 lleaqiasgg vvdpvnsvfl pkdvalargl idrdlyrsln dprdsqknfv dpvtkkkvsy 1741 vqlkercrie phtgllllsv qkrsmsfqgi rqpvtvtelv dsgilrpstv nelesgqisy 1801 devgerikdf lqgssciagi ynettkqklg iyeamkiglv rpgtalelle aqaatgfivd 1861 pvsnlrlpve eaykrglvgi efkekllsae ravtgyndpe tgniislfqa mnkeliekgh 1921 girlleaqia tggiidpkes hrlpvdiayk rgyfneelse ilsdpsddtk gffdpnteen 1981 ltylqlkerc ikdeetglcl lplkekkkqv qtsqkntlrk rrvvivdpet nkemsvqeay 2041 kkglidyetf kelceqecew eeititgsdg strvvlvdrk tgsqydiqda idkglvdrkf 2101 fdqyrsgsls ltqfadmisl kngvgtsssm gsgvsddvfs ssrhesvski stissvrnlt 2161 irsssfsdtl eesspiaaif dtenlekisi tegiergivd sitgqrllea qactggiihp 2221 ttgqklslqd aysqgvidqd matrlkpaqk afigfegvkg kkkmsaaeav kekwlpyeag 2281 qrflefqylt gglvdpevhg risteeairk gfidgraaqr lqdtssyaki ltcpktklki 2341 sykdainrsm veditglrll eaasysskgl pspynmssap gsrsgsrsgs rsgsrsgsrs 2401 gsrrgsfdat gnssysysys fssssigh

In some embodiments of the methods of the disclosure, the wild type human AZGP1 gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_001185.3):

(SEQ ID NO: 21)    1 ccattggcct gtagattcac ctcccctggg cagggcccca ggacccagga taatatctgt   61 gcctcctgcc cagaaccctc caagcagaca caatggtaag aatggtgcct gtcctgctgt  121 ctctgctgct gcttctgggt cctgctgtcc cccaggagaa ccaagatggt cgttactctc  181 tgacctatat ctacactggg ctgtccaagc atgttgaaga cgtccccgcg tttcaggccc  241 ttggctcact caatgacctc cagttcttta gatacaacag taaagacagg aagtctcagc  301 ccatgggact ctggagacag gtggaaggaa tggaggattg gaagcaggac agccaacttc  361 agaaggccag ggaggacatc tttatggaga ccctgaaaga catcgtggag tattacaacg  421 acagtaacgg gtctcacgta ttgcagggaa ggtttggttg tgagatcgag aataacagaa  481 gcagcggagc attctggaaa tattactatg atggaaagga ctacattgaa ttcaacaaag  541 aaatcccagc ctgggtcccc ttcgacccag cagcccagat aaccaagcag aagtgggagg  601 cagaaccagt ctacgtgcag cgggccaagg cttacctgga ggaggagtgc cctgcgactc  661 tgcggaaata cctgaaatac agcaaaaata tcctggaccg gcaagatcct ccctctgtgg  721 tggtcaccag ccaccaggcc ccaggagaaa agaagaaact gaagtgcctg gcctacgact  781 tctacccagg gaaaattgat gtgcactgga ctcgggccgg cgaggtgcag gagcctgagt  841 tacggggaga tgttcttcac aatggaaatg gcacttacca gtcctgggtg gtggtggcag  901 tgcccccgca ggacacagcc ccctactcct gccacgtgca gcacagcagc ctggcccagc  961 ccctcgtggt gccctgggag gccagctagg aagcaagggt tggaggcaat gtgggatctc 1021 agacccagta gctgcccttc ctgcctgatg tgggagctga accacagaaa tcacagtcaa 1081 tggatccaca aggcctgagg agcagtgtgg ggggacagac aggaggtgga tttggagacc 1141 gaagactggg atgcctgtct tgagtagact tggacccaaa aaatcatctc accttgagcc 1201 cacccccacc ccattgtcta atctgtagaa gctaataaat aatcatccct ccttgcctag 1261 cataaaaaaa aaaaaaaa

In some embodiments of the methods of the disclosure, the wild type human AZGP1 gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NP_001176.1):

(SEQ ID NO: 22)   1 mvrmvpvlls lllllgpavp qenqdgrysl tyiytglskh vedvpafqal gslndlqffr  61 ynskdrksqp mglwrqvegm edwkqdsqlq karedifmet lkdiveyynd sngshvlqgr 121 fgceiennrs sgafwkyyyd gkdyiefnke ipawvpfdpa aqitkqkwea epvyvqraka 181 yleeecpatl rkylkyskni ldrqdppsvv vtshqapgek kklkclaydf ypgkidvhwt 241 ragevqepel rgdvlhngng tyqswvvvav ppqdtapysc hvqhsslaqp lvvpweas

In some embodiments of the methods of the disclosure, the wild type human OBFC1 gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_024928):

(SEQ ID NO: 23)    1 aaatgcgctg gcggggagac cggggttggt ccctggcggg gcagggggcg ggctcaggcc   61 ggaactccag agacgacctc agccaactgc tcctgcgccg ggcggggtcg tcgccgccag  121 cggctccgag cgccggaagg gccaggtctc agggctcctg gagctgcagg cggcgggagg  181 ggctacaaat gcttgactca gtgatgcaga acctttcaga gttagctgga agccacagcc  241 ctgcctcttg atgcagcctg gatccagccg gtgtgaagag gagacccctt ccctcttgtg  301 gggtttggat cctgtgtttc tagcctttgc aaaactctac atcagggata tcctggacat  361 gaaggagtcc cgccaggtgc caggtgtatt tttgtacaat ggacatccaa taaaacaggt  421 agatgtcttg ggaactgtca ttggagtgag agaaagagat gctttctaca gttatggagt  481 ggatgacagc actggagtta taaactgcat ctgctggaaa aagttgaata ctgagtctgt  541 atcagctgct ccaagtgcag caagagagct cagcttaacc tcacaactta agaagctaca  601 agagaccatt gagcagaaaa caaagataga gatcggggac acgatccgag tcagaggcag  661 tatccgcaca tacagagaag agcgagagat tcatgccacc acttactata aagtggacga  721 cccagtgtgg aacattcaaa ttgcaaggat gcttgagctg cccactatct acaggaaagt  781 ttatgaccag ccttttcaca gctcagccct agagaaagaa gaggcactaa gcaatccagg  841 cgccctggac ctccccagtc tcacgagttt gctgagtgaa aaagccaaag aattcctcat  901 ggagaacaga gtgcagagct tttaccagca ggagctggaa atggtggagt ctttgctgtc  961 ccttgccaat cagcctgtga ttcacagtgc ctcctccgac caagtgaatt ttaagaagga 1021 caccacttcc aaggcaattc atagtatatt taagaatgct atacaactgc tgcaggaaaa 1081 aggacttgtt ttccagaaag atgatggttt tgataaccta tactatgtaa ccagagaaga 1141 caaagacctg cacagaaaga tccaccggat cattcagcag gactgccaga aaccaaatca 1201 catggagaag ggctgtcact tcctgcacat cttggcctgt gctcgcctga gcatccgccc 1261 gggcctgagc gaggctgtgc tgcagcaagt tctggagctc ctggaggacc agagtgacat 1321 tgtcagcaca atggagcact actacacagc gttctgagca gagacacgca gaccagctga 1381 ggaggacaaa gataaggtgg cattcacccc caggctctga ctttcagcat catgcagggg 1441 cttatctgtc tggaggcagt tacctcataa taaactataa aatatagtca tcttgggaat 1501 gggatttggc ataaatgttg ttggctccct tctgtccact atgtccttgg tgtacaatga 1561 ctttgatctc agccatgaca caacaagaaa accctccctg ttgagctcct ggctggactg 1621 tgcgttgttc gcagagcaga atggggagga aacagtgttg gcagcttaac tgatgtgtgt 1681 ggttggagtc tcttccatgg caaagggaca ccacagggta gtgaacattc aggaactgag 1741 gggcatatgg cctgatcaca cagttctaag cttttcaaaa cttcaggtta tcagagacct 1801 tcctgtgggc ctctcttgct ggctaagaac cggtttaggg gagtagttct ccctggatga 1861 gtgcttacag tttctgtggc tcagttacca gcagtggggt tgagacctgg gtcgatgctc 1921 tttacaggcc tgcccagaga tgggaataaa cagggatcca cagcgtgact atgtgtttgt 1981 cattttcctt ttatttcctt gggaatcgaa aggtgtccca gtacatttcc ctgcacttac 2041 agaggtgcat gactaaatac attgtccctc gatgcccctg aagatcacgg aggcagtcag 2101 ccaattgcct ggcaggtggt agatgttatt ttcagggttg ccgctgagtg tgcaggatgt 2161 gctgacacca tccagacaaa gactcggtat gtgcccagac aggtgatgga gtcatgcttt 2221 tgctcagaat gacaaggtaa aggaaaaaca tctgaggtat gttgtaggcc tgttctgaca 2281 gcaaaatgac aaatccagcc agcaaaaata aagtgtggag aaagatttgg agttaattac 2341 agtcatttca cagaaggcac tgccttcgtc tgctgcattt gctcttgatg tgataagctc 2401 ttcgtggctc agctggagat cctttaggcc tggagagttg ctcctctctc cgtggaaaca 2461 ggacagtctt tatacgcaga agtccgctgc agctcgatac gtcaggctga gagctagaac 2521 cagtagattg cctcctgtca tagacttttg taatgatgca aacctttgct gatttctaac 2581 agtgattatg tagtggctgc cctgcatctt ctctgtgtac agaagggtcc ctagcataga 2641 gtctgcctgg aatgatgtcc tgggcagttc ttccttgagg tcagcagctg ttccacgttg 2701 aatgcatctg attagtgggg ctgcccagga aggagttcag aatcagaagg taaaaagggc 2761 atacccttgc ctatagcaac tctgctctta ggggtttatc tcaaggagat ggctacacaa 2821 gtgtgaaagg atggttgcac aaggtgttca ttgctgtata atctagaatt ctatattggg 2881 gaaaatacct atagggaaaa agttaattac ggttcttggg cacaatgaaa tactatgcag 2941 ctatgaaaaa aatgatgaaa gcagacagac agtgttgcca tggcacactg tccctagtag 3001 atttagtggg aagtagatag agttatagat ctgtttctat agtataacac cattatctac 3061 agctccctgt gtgtatgtat atatccgtag agagagtgta tatttctgca tggaggtctt 3121 tataaatgta gcacatgtac atatatatat atatacacac acacagtcga ccactccctt 3181 ctcctggaag tactttccgc gtttggcttt caggacacca agctctctgg ttgctccttc 3241 tcaggttcct ttgttcagtg ctctgcctcc ctgaggactc agtcccagac ctcttttcta 3301 tctggcttgc tcactggggt gtctccagca gccacatgga ttataccatc tacatgctgt 3361 ctaacacctc agtttaaacc cagaatgggc ctcttccctg aactgcagac ccctatattc 3421 agtttgctac tgacatctcc acttaggtct ctaatggaca tctcagattt cacaggccca 3481 aagccaggct cccaattact cctgacccca ggcttgctcc tgatagtgac atgaggcagc 3541 caaatgccta ggcagagagg ggagggtccc aaatgaaacc ccacgttcaa gcaaagatca 3601 gcctgaaggc taaaagacca gattgctggt cctggatgaa acccaccacg cagagtggga 3661 acttctgttc ctgtttgccc accctttccc aattgttctt tctgaataac gccttaacca 3721 atcgaatgtt gccttttcca gtaataccta cagcctgccc ctccccccat tctgagccca 3781 taaaaagacc cagactcccc catattaagg ggactttcct gcctttgggt agggggacca 3841 cccccacgtc tcctctctgt tgaaaactgt ttcatcactc aataaaactc ccagctttgc 3901 tcactcttcc actgtcagca cattctcatt cttctttggt gctgggcaag aactcaacca 3961 gtgtggaagc catacttggc ccaggcgggt gaagtgggcg ggccgtctcc tgcagcaggt 4021 agcatggtca agcgaggccc aggtgggccg tcaccagcca gaggtccctg gcttgcaaag 4081 tgaccgagaa aaaaatcctg tgccactcct ttggaaaatg tccctgattc aggaagaggt 4141 agctccatcc agttgctcaa accaaatcca ttggcttctt tctttctatc atacctcaca 4201 tccaatctgt ctgcaagtct tttggctcta ccttcagaat atctccagaa tcttaactgc 4261 ttcaccctcc tccccggcct cctcagtcct ctctgcttcc gccctggccc ctcttgggct 4321 gttcacagca cagcagctgt tgccaccctg ttaatgctcc cactctccta cagccttcgg 4381 tcttgcccca ggtaggagcc tgaggctgca cagaggtcag cacggccccg cttaccctgc 4441 cctcccagcc cagccgcacg ggccttgcac acatgcctcg gcatattcct gccttagggc 4501 tggtgctcct gctatttcct cttcccaggt aaccatgtga agtgcctccc tctgccctct 4561 ttccagcctt tacttgagtg tcaccttctc agtgaggcct gccctcattc ctctttcgct 4621 gtttgcaacc catctcctgt cccccttccc agaactccct ttcctacttc gtttttcttc 4681 acagtacttg atactgccta acacactcca tggtttctta cttgccctgt ttattatttt 4741 cccccaatag acagaatgtt ccatgatggc agaattctct gttttgtttc cttccatgtc 4801 cccagcacct agaacagtgc ctgacgcatc tcctaagcaa tacgaccaat aagtatgtgt 4861 ctggctgcct tccggctgcc agtgtctgcc tctttcctag gggcagtggt tgcgggggtg 4921 ctttctcaca tgtcttagta ggctgtgcag gctggaagtg ctcagaagtc acacccccag 4981 ggagcagcct cagccaacag caccttggct gtaaatgccc cagctccctc gccctcaggt 5041 aagcattgct gaggcacacg ttccatactc ttttccacag ttcctccgtg ggactgagca 5101 ccacccagcc acccacagga gcagctaacc tgataaccac cagcctcacc ctccctgcct 5161 tacttccccg ctccccttta ccacatgctg acctcccaga tgcatttctt gctttccggt 5221 ctctgtctca ggattggctc ctggatgaac acaaactaac actatgttca caaatatatt 5281 tgggaaatgc tggatgaata attatacaca tcagacagat tactagaaat tctcaccaaa 5341 gggatgcaca tgttacctct gcatggtgag atctcaggtg ctttttaccc cacatagcta 5401 tcctttggca tttttataat tagcaagtgc tcactcttcc actgtcagta cattctcatt 5461 cttcttgggc gctggacaag aattcaaccg gtgtgtaagc cagactcggc ccgggcagtc 5521 tcaaactcct gactccttat ataatttcta caaaaattat aaagctattt cccactcccc 5581 accccacatt catgtaacct gaagcatgag taaaccaaga atgaggtagg cctctgtctt 5641 ctaagcaaca tcagaactct aagaacatga gggactctta gaaaactctc tggagctaac 5701 cacagctggg tcactgctca tgtactgaag accagccaga gggttcccct gaaaaggagg 5761 gaaactgagc aaacattctc cagttctctt agtgtgcaca tgtttcagga ggtgtgaacc 5821 ccacatgtag cttgtgtagg caagaagaca aatagtgcta ctgtctggtc aaggatttgt 5881 ttgaagagcc atgattatgc ccatatggta agccaccagt gctccccatc cctgtaagac 5941 acttctttct cattattttc tcctctgatg gtgtgccagg atgctggcca agagaagcca 6001 agtggaaaga aggctgttca gtgacaagga acctaagact tagtgccaag gactgaaacc 6061 aagtaaactt gtaattttcc atgatggaaa catctacact ttctcattag tggcctctac 6121 agcagttgcc ccaaagaagc gtctcattgt ttttttacta catttatgtg aagcatacag 6181 gcaaactcag aaagactgtg ataaggctcg ccagagatgc ctgcacaggt gctgggggaa 6241 aagcaggacc atcctgaagg gagatggtgt ctgtggacaa agaactctgc agtggttctt 6301 atttgcatga tttctgctgg tggaggctgt aaatgtgagc tcaaactccc acataagtga 6361 gttttcattg taatccagaa tgtttttaaa tcaccctact tctattgaac ttgcactatc 6421 atctgttaac ctctactgta tttattaaat aaacctgaat aggtaaatca cagtacagca 6481 aaa

In some embodiments of the methods of the disclosure, the wild type human OBFC1 gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_079204.2):

(SEQ ID NO: 24)   1 mqpgssrcee etpsllwgld pvflafakly irdildmkes rqvpgvflyn ghpikqvdvl  61 gtvigvrerd afysygvdds tgvincicwk klntesvsaa psaarelslt sqlkklqeti 121 eqktkieigd tirvrgsirt yreereihat tyykvddpvw niqiarmlel ptiyrkvydq 181 pfhssaleke ealsnpgald lpsltsllse kakeflmenr vqsfyqqele mvesllslan 241 qpvihsassd qvnfkkdtts kaihsifkna iqllqekglv fqkddgfdnl yyvtredkdl 301 hrkihriiqq dcqkpnhmek gchflhilac arlsirpgls eavlqqvlel ledqsdivst 361 mehyytaf

In some embodiments of the methods of the disclosure, the wild type human ATP11A gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_015205.2, transcript variant 1):

(SEQ ID NO: 25)    1 gcggccgcac tagtaccccg gagcccatgg gcgcgccgag ccgggcgcgg gggcgctgaa   61 cggcggagcg ggagcggccg gaggagccat ggactgcagc ctcgtgcgga cgctcgtgca  121 cagatactgt gcaggagaag agaattgggt ggacagcagg accatctacg tgggacacag  181 ggagccacct ccgggcgcag aggcctacat cccacagaga tacccagaca acaggatcgt  241 ctcgtccaag tacacatttt ggaactttat acccaagaat ttatttgaac aattcagaag  301 agtagccaac ttttatttcc ttatcatatt tctggtgcag ttgattattg atacacccac  361 aagtccagtg acaagcggac ttccactctt ctttgtcatt actgtgacgg ctatcaaaca  421 gggttatgaa gactggcttc gacataaagc agacaatgcc atgaaccagt gtcctgttca  481 tttcattcag cacggcaagc tcgttcggaa acaaagtcga aagctgcgag ttggggacat  541 tgtcatggtt aaggaggacg agacctttcc ctgcgacttg atcttccttt ccagcaaccg  601 gggagatggg acgtgccacg tcaccaccgc cagcttggat ggagaatcca gccataaaac  661 gcattacgcg gtccaggaca ccaaaggctt ccacacagag gaggatatcg gcggacttca  721 cgccaccatc gagtgtgagc agccccagcc cgacctctac aagttcgtgg gtcgcatcaa  781 cgtttacagt gacctgaatg accccgtggt gaggccctta ggatcggaaa acctgctgct  841 tagaggagct acactgaaga acactgagaa aatctttggt gtggctattt acacgggaat  901 ggaaaccaag atggcattaa attatcaatc aaaatctcag aagcgatctg ccgtggaaaa  961 atcgatgaat gcgttcctca ttgtgtatct ctgcattctg atcagcaaag ccctgataaa 1021 cactgtgctg aaatacatgt ggcagagtga gccctttcgg gatgagccgt ggtataatca 1081 gaaaacggag tcggaaaggc agaggaatct gttcctcaag gcattcacgg acttcctggc 1141 cttcatggtc ctctttaact acatcatccc tgtgtccatg tacgtcacgg tcgagatgca 1201 gaagttcctc ggctcttact tcatcacctg ggacgaagac atgtttgacg aggagactgg 1261 cgaggggcct ctggtgaaca cgtcggacct caatgaagag ctgggacagg tggagtacat 1321 cttcacagac aagaccggca ccctcacgga aaacaacatg gagttcaagg agtgctgcat 1381 cgaaggccat gtctacgtgc cccacgtcat ctgcaacggg caggtcctcc cagagtcgtc 1441 aggaatcgac atgattgact cgtcccccag cgtcaacggg agggagcgcg aggagctgtt 1501 tttccgggcc ctctgtctct gccacaccgt ccaggtgaaa gacgatgaca gcgtagacgg 1561 ccccaggaaa tcgccggacg gggggaaatc ctgtgtgtac atctcatcct cgcccgacga 1621 ggtggcgctg gtcgaaggtg tccagagact tggctttacc tacctaaggc tgaaggacaa 1681 ttacatggag atattaaaca gggagaacca catcgaaagg tttgaattgc tggaaatttt 1741 gagttttgac tcagtcagaa ggagaatgag tgtaattgta aaatctgcta caggagaaat 1801 ttatctgttt tgcaaaggag cagattcttc gatattcccc cgagtgatag aaggcaaagt 1861 tgaccagatc cgagccagag tggagcgtaa cgcagtggag gggctccgaa ctttgtgtgt 1921 tgcttataaa aggctgatcc aagaagaata tgaaggcatt tgtaagctgc tgcaggctgc 1981 caaagtggcc cttcaagatc gagagaaaaa gttagcagaa gcctatgagc aaatagagaa 2041 agatcttact ctgcttggtg ctacagctgt tgaggaccgg ctgcaggaga aagctgcaga 2101 caccatcgag gccctgcaga aggccgggat caaagtctgg gttctcacgg gagacaagat 2161 ggagacggcc gcggccacgt gctacgcctg caagctcttc cgcaggaaca cgcagctgct 2221 ggagctgacc accaagagga tcgaggagca gagcctgcac gacgtcctgt tcgagctgag 2281 caagacggtc ctgcgccaca gcgggagcct gaccagagac aacctgtccg gactttcagc 2341 agatatgcag gactacggtt taattatcga cggagctgca ctgtctctga taatgaagcc 2401 tcgagaagac gggagttccg gcaactacag ggagctcttc ctggaaatct gccggagctg 2461 cagcgcggtg ctctgctgcc gcatggcgcc cttgcagaag gctcagattg ttaaattaat 2521 caaattttca aaagagcacc caatcacgtt agcaattggc gatggtgcaa atgatgtcag 2581 catgattctg gaagcgcacg tgggcatagg tgtcatcggc aaggaaggcc gccaggctgc 2641 caggaacagc gactatgcaa tcccaaagtt taagcatttg aagaagatgc tgcttgttca 2701 cgggcatttt tattacatta ggatctctga gctcgtgcag tacttcttct ataagaacgt 2761 ctgcttcatc ttccctcagt ttttatacca gttcttctgt gggttttcac aacagacttt 2821 gtacgacacc gcgtatctga ccctctacaa catcagcttc acctccctcc ccatcctcct 2881 gtacagcctc atggagcagc atgttggcat tgacgtgctc aagagagacc cgaccctgta 2941 cagggacgtc gccaagaatg ccctgctgcg ctggcgcgtg ttcatctact ggacgctcct 3001 gggactgttt gacgcactgg tgttcttctt tggtgcttat ttcgtgtttg aaaatacaac 3061 tgtgacaagc aacgggcaga tatttggaaa ctggacgttt ggaacgctgg tattcaccgt 3121 gatggtgttc acagttacac taaagcttgc attggacaca cactactgga cttggatcaa 3181 ccattttgtc atctgggggt cgctgctgtt ctacgttgtc ttttcgcttc tctggggagg 3241 agtgatctgg ccgttcctca actaccagag gatgtactac gtgttcatcc agatgctgtc 3301 cagcgggccc gcctggctgg ccatcgtgct gctggtgacc atcagcctcc ttcccgacgt 3361 cctcaagaaa gtcctgtgcc ggcagctgtg gccaacagca acagagagag tccagactaa 3421 gagccagtgc ctttctgtcg agcagtcaac catctttatg ctttctcaga cttccagcag 3481 cctgagtttc tgatggaaca agagcccagg ctaccagagc acctgtccct cggccgcctg 3541 gtacagctcc cactctcagc aggtgacact cgcggcctgg aaggagaagg tgtccacgga 3601 gcccccaccc atcctcggcg gttcccatca ccactgcagt tccatcccaa gtcacagctg 3661 ccctaggtcc cgtgtgggaa tgctcgtgtg atggatggtc ctaagcctgt ggagactgtg 3721 cacgtgcctc ttcctggccc ccagcaggca aggagggggg tcacaggcct tgccctcgag 3781 catggcaccc tggccgcctg gacccagcac tgtggttgtt gagccacacc agtggcctct 3841 gggcattcgg ctcaacgcag gagggacatt ctgctggccc accctgcgcg ctgtcatgca 3901 gaggccattc ccccaggcct gtgtcttcac ccacctgcca tcattggcct ttgctgtcac 3961 tgggagagaa gagccgtcca gggacccatg gtggcccaca tgtggatgcc acatgctgct 4021 gtttcctgct tgcccggcca ccacccatgc cctccatagg gtgaggtgga gccatggtgg 4081 tgcgtccttt actcaacaac cctccaatcc ggatgctgtg ggaagggccg ggtcactcgg 4141 ataccatcat ccctgcggat gcaccgccgt accctgctca tctgggagtg gtttccctgc 4201 ggttacgtcc aagcccgcct gccctgtgtg ttggggctgg ctgagtttcg gtctccccat 4261 caccggccgc ctcgtggaga aggcagtgcc acgtgggagg acaaggccac gccggcagct 4321 tccagccctg ccgcagaagt gccaggatgt ccatcagcca ctcgccaggg cacggagccg 4381 tcagtccact gttacgggag aatgttgatt tcgcgggtgc gagggccggg agacagatac 4441 ttggctgtga tgagcagaca tcctctgtcc ccgtggaggg gtcaacacca aggtggtgtt 4501 cgtgcaccag aacctgtctc gggctgacgg gggtggcaca caggacacgg gtggatccca 4561 acaggcagca ccgcacctct gcccgcctcc cgcactgcag ctccgcccgc cgggctctgc 4621 gtccccacgt cccctcgtcc catccccacg tcccctcatc ccgtcacctc gtccccacat 4681 ccccttgccc cgtcacctcg tcctcatgtc cccttgtcct gtcacctcgt ccccacgtcc 4741 cctcgtctcc tcatccccac gtcctctcgt ccccttgtcc cgtccccaca taccctcgtc 4801 cccatgtccc cacgcagggc tctccttcgt cttaggatct gtccagcgct gctctgggtg 4861 ggttagcaac cccagggctg ctgtgatagg aagtccctgt tgttctccgt actggcattt 4921 ctatttctag aaataatatt tgacatagcc ttaatggtcc ttaaagaaga catttcagtg 4981 tgagattcag acttcagacg ctgaaactgc tgcctttcag gaaagcacca ccaacgctgg 5041 aggaggagcc ggccctcacg cccgccccgc gccacgctgt ggaacggggc tccggcaagt 5101 gaaacccaga gggtgtttcc gaggtgctcg acagtaggta tttttggaag ctcagatttc 5161 accatttgat tgtataatct tttacctata aaatatttat ttgaagtaga gggtaaatca 5221 gcggtaagaa cagtgaacac agtggttggg ataaaataag gtgacaaaca tcacaccaaa 5281 gatgagggta gcgagcaact ggcttgagca gacagaacgg ggaagactcc actctgtccc 5341 gaggggccag ccgcaggcgt ccccagggcc accctgccct gaggtccttg tgtggccgcc 5401 ctggcttggc agccctgccc acgctgcccc cgcaaacaat ggtgtgtgcg tttttacagc 5461 cctttttagg aacccaatat gggcataaat gtaacacctg tagcgggggc agattctctg 5521 tatgttcagt taacaaatta tttgtaatgt atttttttag aaatcttaaa attgcctttg 5581 cactgaagta ttttcatagc tgtttatatc tcttttattc atttatttaa catactgtct 5641 aattttaaaa ataggttttt aaagctttca tttttaagtt tatgaaattt tggccacttt 5701 acatttagat tctggtgaga gttttgactg aatgttccaa tctctgatga atgcgaattt 5761 tcagatttga ttttattctc tacacacacc tcttcttttc ttggtatttc tggtggcagt 5821 gattagttga acagcacatt taaggcacga taatttgcta cactttttct ttacaatttg 5881 ttgcaatttc atctgctttc tatgtttcat tgttaattgc catccttcag ccttaaaaat 5941 agaagattct cacgtgaagg tttagtaagt tgggtcccag ctctgcctgt gtggagatag 6001 tcaccatgta cctctgacaa caagttttag tgtgaaagtc actaaacttt tacacactcc 6061 caaacgtctt tttaaaaatt gcttgggaaa ttattaaatg aatgtgcctg atgatttgaa 6121 atagacaagg ggcacgagat aaaaaagaaa aggatgagaa gatcctcagt gaatgacgtt 6181 gcagggtctt catgcaattt tccacctcgc agtagttagt atttacttgc cttaaactaa 6241 ctttgaagca agtaatgtca actttgagca ctttgttgag ttttgaaaaa tcttatttgt 6301 tgctgcacag gttaataaat tatcaatttg taattcagca tgttggtcag agacacggtc 6361 actgattcac acccagtccc tgccacagac cgtctcagac acgcacagtg ggcctgctgc 6421 atgattcaca cccagtccct gccacagacc gtctcagaca cgcacagtgg gcctgctgca 6481 tgattcacac ccagtccctg ccacagaccg tctcagacac gcacagtggg cctgctgcat 6541 gcgtgttacc tggcttttgg ctccacgctc actcatagcc atgtccacat gggggcttgc 6601 acacaggatc actcacatat gtacatgtac ccaccacaaa cgtgcaagct cctgcacaca 6661 tgcatgcaca caaacgtgta cacaagtgtg agctcctaca cgcatacaca cacacacgtg 6721 tacatgcacc aaagcatgtg tgacctacag acatgcagaa catgcacgtg tacacatacc 6781 acagacacgc gtgtgcatgc tcctacacaa tacatatgca catatcatga acagcgtaag 6841 ttcctacaca cggacgtgtg atacacacat gcatgtacag gtaagcacac atgtacaagc 6901 tcctacaggc ttgctctcac acacgtgtat gcacagcaga gagacgtatg agcttctact 6961 gcacacatgc acacacacac gcacacgtac attcactaca aacgtgcagc ctcctgcaca 7021 cgtgcacatt catgtgtaca ccacaaatga gttcccagac gtgtaaacac acgtgcacac 7081 atcgtacaca tgtgagctcc cacacgtaca cacagatgca catggacaca ccccaaacac 7141 gcacaggctc ctacacacat gcacacacgt gtacaccaca aacgagctcc cagacatgta 7201 aacacacgtc tcccacacgt gagctcccac acgtacacat gcacatgtac gcaccacaaa 7261 cacatgcgca ggctcctgca ggcgtgaata cacacatgca cacacatata cacacatgtg 7321 ccacaaacaa gtgcacactg tcctggtgtc ctgcactgca tcctgcctcc ttgctgaggg 7381 gcccctgtga gaggcctctg gatgggcatg ggaagatggg ctccctggcc cccagcccat 7441 gcctccctgg gatgaagagt ccccctcctg gcagaatgtc tgggctttgc agagcaggcc 7501 ccgggggtga agtcgcagct tcacttacac cagctgctct gtgagcaagg cttggtgccc 7561 tggacaaggc ccttcccctt tagggaggtc cagcctcgca agctgaaacc tcccctcggc 7621 tcagccctat accaggcggc cacagcagga ctggccacac ccacgccgca cctcatccgt 7681 gcacgcgtcg gagcacggcc agccttccgc cacgagccag ctgggaaggg ccgcggccgc 7741 ctaaagcccc agtcaaccca gcctgtgtct gagcagacag ggcgaacaag caggccacac 7801 cgtctcgagg gaggaggcca gatgcggcca gcgtctccaa cagggtgacc atccgctcgg 7861 cttgctgagc gtttaaacaa atgtttagac aggctgtggg gactcccctg agttgagcct 7921 tggccagggg tccggtgctg tcgcgggaaa cctccagcct tgttcttcaa accactcagc 7981 tcatgtgttt tgcactgact agtactgaat aatacaacca ctcttattta atgttagtat 8041 tatttatttg acaactcagt gtctaacagc ttgatatgca ggtccttgca tcctacattt 8101 ctttaggaag ttacccattt gtaactttaa aaacaggaaa aatatcagtt ggcaaatgca 8161 atcttttttt tttttaagct aaaggtgggt gaactggaat gaaaatcttt ctgatgttgt 8221 gtctataagc agccttgatg ggatatgtta gaagtgtcat gaaagtgtga ttctactttt 8281 gcagaaaaat ctaaagatca atttatatag ctttattttt tactttatca aagtatacag 8341 aattttaata tgcatatatt gtgtctgact taaaattata atgtctgcgt caccatttaa 8401 aatgtctgtt cattatgtaa tgtaataaaa gaaggtcttc aaaaatgtat ttaacatgaa 8461 tggtatccat agttgtcatc atcataaata ctggagttta tttttaaatt attaaacata 8521 gtaggtgcat taacataaat cagtctccac acagtaacat ttaactgata attcattaat 8581 cagctttgaa aaattaaatt gttaattaaa ccaatctaac atttcagtaa agtttatttt 8641 gtatgcttct gtttttaact tttatttctg tagataaact gactggataa tattatattg 8701 gacttttctc tagattatct aagcaggaga cctgaatctg cttgcaataa agaataaaag 8761 tctgcttcag tttctttata aagaaactca cacaa

In some embodiments of the methods of the disclosure, the wild type human

ATP11A gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NP_056020.2, transcript variant 1):

(SEQ ID NO: 26)    1 mdcslvrtlv hrycageenw vdsrtiyvgh repppgaeay ipqrypdnri vsskytfwnf   61 ipknlfeqfr rvanfyflii flvqliidtp tspvtsglpl ffvitvtaik qgyedwlrhk  121 adnamnqcpv hfiqhgklvr kqsrklrvgd ivmvkedetf pcdliflssn rgdgtchvtt  181 asldgesshk thyavqdtkg fhteediggl hatieceqpq pdlykfvgri nvysdlndpv  241 vrplgsenll lrgatlknte kifgvaiytg metkmalnyq sksqkrsave ksmnaflivy  301 lciliskali ntvlkymwqs epfrdepwyn qkteserqrn lflkaftdfl afmvlfnyii  361 pvsmyvtvem qkflgsyfit wdedmfdeet gegplvntsd lneelgqvey iftdktgtlt  421 ennmefkecc ieghvyvphv icngqvlpes sgidmidssp svngrereel ffralclcht  481 vqvkdddsvd gprkspdggk scvyissspd evalvegvqr lgftylrlkd nymeilnren  541 hierfellei lsfdsvrrrm svivksatge iylfckgads sifprviegk vdqirarver  601 naveglrtlc vaykrliqee yegickllqa akvalqdrek klaeayeqie kdltllgata  661 vedrlqekaa dtiealqkag ikvwvltgdk metaaatcya cklfrrntql lelttkriee  721 qslhdvlfel sktvlrhsgs ltrdnlsgls admqdyglii dgaalslimk predgssgny  781 relfleicrs csavlccrma plqkaqivkl ikfskehpit laigdgandv smileahvgi  841 gvigkegrqa arnsdyaipk fkhlkkmllv hghfyyiris elvqyffykn vcfifpqfly  901 qffcgfsqqt lydtayltly nisftslpil lyslmeqhvg idvlkrdptl yrdvaknall  961 rwrvfiywtl lglfdalvff fgayfvfent tvtsngqifg nwtfgtlvft vmvftvtlkl 1021 aldthywtwi nhfviwgsll fyvvfsllwg gviwpflnyq rmyyvfiqml ssgpawlaiv 1081 llvtisllpd vlkkvlcrql wptatervqt ksqclsveqs tifmlsqtss slsf

In some embodiments of the methods of the disclosure, the wild type human ATP11A gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_032189.3, transcript variant 2):

(SEQ ID NO: 48)    1 gcggccgcac tagtaccccg gagcccatgg gcgcgccgag ccgggcgcgg gggcgctgaa   61 cggcggagcg ggagcggccg gaggagccat ggactgcagc ctcgtgcgga cgctcgtgca  121 cagatactgt gcaggagaag agaattgggt ggacagcagg accatctacg tgggacacag  181 ggagccacct ccgggcgcag aggcctacat cccacagaga tacccagaca acaggatcgt  241 ctcgtccaag tacacatttt ggaactttat acccaagaat ttatttgaac aattcagaag  301 agtagccaac ttttatttcc ttatcatatt tctggtgcag ttgattattg atacacccac  361 aagtccagtg acaagcggac ttccactctt ctttgtcatt actgtgacgg ctatcaaaca  421 gggttatgaa gactggcttc gacataaagc agacaatgcc atgaaccagt gtcctgttca  481 tttcattcag cacggcaagc tcgttcggaa acaaagtcga aagctgcgag ttggggacat  541 tgtcatggtt aaggaggacg agacctttcc ctgcgacttg atcttccttt ccagcaaccg  601 gggagatggg acgtgccacg tcaccaccgc cagcttggat ggagaatcca gccataaaac  661 gcattacgcg gtccaggaca ccaaaggctt ccacacagag gaggatatcg gcggacttca  721 cgccaccatc gagtgtgagc agccccagcc cgacctctac aagttcgtgg gtcgcatcaa  781 cgtttacagt gacctgaatg accccgtggt gaggccctta ggatcggaaa acctgctgct  841 tagaggagct acactgaaga acactgagaa aatctttggt gtggctattt acacgggaat  901 ggaaaccaag atggcattaa attatcaatc aaaatctcag aagcgatctg ccgtggaaaa  961 atcgatgaat gcgttcctca ttgtgtatct ctgcattctg atcagcaaag ccctgataaa 1021 cactgtgctg aaatacatgt ggcagagtga gccctttcgg gatgagccgt ggtataatca 1081 gaaaacggag tcggaaaggc agaggaatct gttcctcaag gcattcacgg acttcctggc 1141 cttcatggtc ctctttaact acatcatccc tgtgtccatg tacgtcacgg tcgagatgca 1201 gaagttcctc ggctcttact tcatcacctg ggacgaagac atgtttgacg aggagactgg 1261 cgaggggcct ctggtgaaca cgtcggacct caatgaagag ctgggacagg tggagtacat 1321 cttcacagac aagaccggca ccctcacgga aaacaacatg gagttcaagg agtgctgcat 1381 cgaaggccat gtctacgtgc cccacgtcat ctgcaacggg caggtcctcc cagagtcgtc 1441 aggaatcgac atgattgact cgtcccccag cgtcaacggg agggagcgcg aggagctgtt 1501 tttccgggcc ctctgtctct gccacaccgt ccaggtgaaa gacgatgaca gcgtagacgg 1561 ccccaggaaa tcgccggacg gggggaaatc ctgtgtgtac atctcatcct cgcccgacga 1621 ggtggcgctg gtcgaaggtg tccagagact tggctttacc tacctaaggc tgaaggacaa 1681 ttacatggag atattaaaca gggagaacca catcgaaagg tttgaattgc tggaaatttt 1741 gagttttgac tcagtcagaa ggagaatgag tgtaattgta aaatctgcta caggagaaat 1801 ttatctgttt tgcaaaggag cagattcttc gatattcccc cgagtgatag aaggcaaagt 1861 tgaccagatc cgagccagag tggagcgtaa cgcagtggag gggctccgaa ctttgtgtgt 1921 tgcttataaa aggctgatcc aagaagaata tgaaggcatt tgtaagctgc tgcaggctgc 1981 caaagtggcc cttcaagatc gagagaaaaa gttagcagaa gcctatgagc aaatagagaa 2041 agatcttact ctgcttggtg ctacagctgt tgaggaccgg ctgcaggaga aagctgcaga 2101 caccatcgag gccctgcaga aggccgggat caaagtctgg gttctcacgg gagacaagat 2161 ggagacggcc gcggccacgt gctacgcctg caagctcttc cgcaggaaca cgcagctgct 2221 ggagctgacc accaagagga tcgaggagca gagcctgcac gacgtcctgt tcgagctgag 2281 caagacggtc ctgcgccaca gcgggagcct gaccagagac aacctgtccg gactttcagc 2341 agatatgcag gactacggtt taattatcga cggagctgca ctgtctctga taatgaagcc 2401 tcgagaagac gggagttccg gcaactacag ggagctcttc ctggaaatct gccggagctg 2461 cagcgcggtg ctctgctgcc gcatggcgcc cttgcagaag gctcagattg ttaaattaat 2521 caaattttca aaagagcacc caatcacgtt agcaattggc gatggtgcaa atgatgtcag 2581 catgattctg gaagcgcacg tgggcatagg tgtcatcggc aaggaaggcc gccaggctgc 2641 caggaacagc gactatgcaa tcccaaagtt taagcatttg aagaagatgc tgcttgttca 2701 cgggcatttt tattacatta ggatctctga gctcgtgcag tacttcttct ataagaacgt 2761 ctgcttcatc ttccctcagt ttttatacca gttcttctgt gggttttcac aacagacttt 2821 gtacgacacc gcgtatctga ccctctacaa catcagcttc acctccctcc ccatcctcct 2881 gtacagcctc atggagcagc atgttggcat tgacgtgctc aagagagacc cgaccctgta 2941 cagggacgtc gccaagaatg ccctgctgcg ctggcgcgtg ttcatctact ggacgctcct 3001 gggactgttt gacgcactgg tgttcttctt tggtgcttat ttcgtgtttg aaaatacaac 3061 tgtgacaagc aacgggcaga tatttggaaa ctggacgttt ggaacgctgg tattcaccgt 3121 gatggtgttc acagttacac taaagcttgc attggacaca cactactgga cttggatcaa 3181 ccattttgtc atctgggggt cgctgctgtt ctacgttgtc ttttcgcttc tctggggagg 3241 agtgatctgg ccgttcctca actaccagag gatgtactac gtgttcatcc agatgctgtc 3301 cagcgggccc gcctggctgg ccatcgtgct gctggtgacc atcagcctcc ttcccgacgt 3361 cctcaagaaa gtcctgtgcc ggcagctgtg gccaacagca acagagagag tccagaatgg 3421 gtgcgcacag cctcgggacc gcgactcaga attcacccct cttgcctctc tgcagagccc 3481 aggctaccag agcacctgtc cctcggccgc ctggtacagc tcccactctc agcaggtgac 3541 actcgcggcc tggaaggaga aggtgtccac ggagccccca cccatcctcg gcggttccca 3601 tcaccactgc agttccatcc caagtcacag ctgccctagg tcccgtgtgg gaatgctcgt 3661 gtgatggatg gtcctaagcc tgtggagact gtgcacgtgc ctcttcctgg cccccagcag 3721 gcaaggaggg gggtcacagg ccttgccctc gagcatggca ccctggccgc ctggacccag 3781 cactgtggtt gttgagccac accagtggcc tctgggcatt cggctcaacg caggagggac 3841 attctgctgg cccaccctgc gcgctgtcat gcagaggcca ttcccccagg cctgtgtctt 3901 cacccacctg ccatcattgg cctttgctgt cactgggaga gaagagccgt ccagggaccc 3961 atggtggccc acatgtggat gccacatgct gctgtttcct gcttgcccgg ccaccaccca 4021 tgccctccat agggtgaggt ggagccatgg tggtgcgtcc tttactcaac aaccctccaa 4081 tccggatgct gtgggaaggg ccgggtcact cggataccat catccctgcg gatgcaccgc 4141 cgtaccctgc tcatctggga gtggtttccc tgcggttacg tccaagcccg cctgccctgt 4201 gtgttggggc tggctgagtt tcggtctccc catcaccggc cgcctcgtgg agaaggcagt 4261 gccacgtggg aggacaaggc cacgccggca gcttccagcc ctgccgcaga agtgccagga 4321 tgtccatcag ccactcgcca gggcacggag ccgtcagtcc actgttacgg gagaatgttg 4381 atttcgcggg tgcgagggcc gggagacaga tacttggctg tgatgagcag acatcctctg 4441 tccccgtgga ggggtcaaca ccaaggtggt gttcgtgcac cagaacctgt ctcgggctga 4501 cgggggtggc acacaggaca cgggtggatc ccaacaggca gcaccgcacc tctgcccgcc 4561 tcccgcactg cagctccgcc cgccgggctc tgcgtcccca cgtcccctcg tcccatcccc 4621 acgtcccctc atcccgtcac ctcgtcccca catccccttg ccccgtcacc tcgtcctcat 4681 gtccccttgt cctgtcacct cgtccccacg tcccctcgtc tcctcatccc cacgtcctct 4741 cgtccccttg tcccgtcccc acataccctc gtccccatgt ccccacgcag ggctctcctt 4801 cgtcttagga tctgtccagc gctgctctgg gtgggttagc aaccccaggg ctgctgtgat 4861 aggaagtccc tgttgttctc cgtactggca tttctatttc tagaaataat atttgacata 4921 gccttaatgg tccttaaaga agacatttca gtgtgagatt cagacttcag acgctgaaac 4981 tgctgccttt caggaaagca ccaccaacgc tggaggagga gccggccctc acgcccgccc 5041 cgcgccacgc tgtggaacgg ggctccggca agtgaaaccc agagggtgtt tccgaggtgc 5101 tcgacagtag gtatttttgg aagctcagat ttcaccattt gattgtataa tcttttacct 5161 ataaaatatt tatttgaagt agagggtaaa tcagcggtaa gaacagtgaa cacagtggtt 5221 gggataaaat aaggtgacaa acatcacacc aaagatgagg gtagcgagca actggcttga 5281 gcagacagaa cggggaagac tccactctgt cccgaggggc cagccgcagg cgtccccagg 5341 gccaccctgc cctgaggtcc ttgtgtggcc gccctggctt ggcagccctg cccacgctgc 5401 ccccgcaaac aatggtgtgt gcgtttttac agcccttttt aggaacccaa tatgggcata 5461 aatgtaacac ctgtagcggg ggcagattct ctgtatgttc agttaacaaa ttatttgtaa 5521 tgtatttttt tagaaatctt aaaattgcct ttgcactgaa gtattttcat agctgtttat 5581 atctctttta ttcatttatt taacatactg tctaatttta aaaataggtt tttaaagctt 5641 tcatttttaa gtttatgaaa ttttggccac tttacattta gattctggtg agagttttga 5701 ctgaatgttc caatctctga tgaatgcgaa ttttcagatt tgattttatt ctctacacac 5761 acctcttctt ttcttggtat ttctggtggc agtgattagt tgaacagcac atttaaggca 5821 cgataatttg ctacactttt tctttacaat ttgttgcaat ttcatctgct ttctatgttt 5881 cattgttaat tgccatcctt cagccttaaa aatagaagat tctcacgtga aggtttagta 5941 agttgggtcc cagctctgcc tgtgtggaga tagtcaccat gtacctctga caacaagttt 6001 tagtgtgaaa gtcactaaac ttttacacac tcccaaacgt ctttttaaaa attgcttggg 6061 aaattattaa atgaatgtgc ctgatgattt gaaatagaca aggggcacga gataaaaaag 6121 aaaaggatga gaagatcctc agtgaatgac gttgcagggt cttcatgcaa ttttccacct 6181 cgcagtagtt agtatttact tgccttaaac taactttgaa gcaagtaatg tcaactttga 6241 gcactttgtt gagttttgaa aaatcttatt tgttgctgca caggttaata aattatcaat 6301 ttgtaattca gcatgttggt cagagacacg gtcactgatt cacacccagt ccctgccaca 6361 gaccgtctca gacacgcaca gtgggcctgc tgcatgattc acacccagtc cctgccacag 6421 accgtctcag acacgcacag tgggcctgct gcatgattca cacccagtcc ctgccacaga 6481 ccgtctcaga cacgcacagt gggcctgctg catgcgtgtt acctggcttt tggctccacg 6541 ctcactcata gccatgtcca catgggggct tgcacacagg atcactcaca tatgtacatg 6601 tacccaccac aaacgtgcaa gctcctgcac acatgcatgc acacaaacgt gtacacaagt 6661 gtgagctcct acacgcatac acacacacac gtgtacatgc accaaagcat gtgtgaccta 6721 cagacatgca gaacatgcac gtgtacacat accacagaca cgcgtgtgca tgctcctaca 6781 caatacatat gcacatatca tgaacagcgt aagttcctac acacggacgt gtgatacaca 6841 catgcatgta caggtaagca cacatgtaca agctcctaca ggcttgctct cacacacgtg 6901 tatgcacagc agagagacgt atgagcttct actgcacaca tgcacacaca cacgcacacg 6961 tacattcact acaaacgtgc agcctcctgc acacgtgcac attcatgtgt acaccacaaa 7021 tgagttccca gacgtgtaaa cacacgtgca cacatcgtac acatgtgagc tcccacacgt 7081 acacacagat gcacatggac acaccccaaa cacgcacagg ctcctacaca catgcacaca 7141 cgtgtacacc acaaacgagc tcccagacat gtaaacacac gtctcccaca cgtgagctcc 7201 cacacgtaca catgcacatg tacgcaccac aaacacatgc gcaggctcct gcaggcgtga 7261 atacacacat gcacacacat atacacacat gtgccacaaa caagtgcaca ctgtcctggt 7321 gtcctgcact gcatcctgcc tccttgctga ggggcccctg tgagaggcct ctggatgggc 7381 atgggaagat gggctccctg gcccccagcc catgcctccc tgggatgaag agtccccctc 7441 ctggcagaat gtctgggctt tgcagagcag gccccggggg tgaagtcgca gcttcactta 7501 caccagctgc tctgtgagca aggcttggtg ccctggacaa ggcccttccc ctttagggag 7561 gtccagcctc gcaagctgaa acctcccctc ggctcagccc tataccaggc ggccacagca 7621 ggactggcca cacccacgcc gcacctcatc cgtgcacgcg tcggagcacg gccagccttc 7681 cgccacgagc cagctgggaa gggccgcggc cgcctaaagc cccagtcaac ccagcctgtg 7741 tctgagcaga cagggcgaac aagcaggcca caccgtctcg agggaggagg ccagatgcgg 7801 ccagcgtctc caacagggtg accatccgct cggcttgctg agcgtttaaa caaatgttta 7861 gacaggctgt ggggactccc ctgagttgag ccttggccag gggtccggtg ctgtcgcggg 7921 aaacctccag ccttgttctt caaaccactc agctcatgtg ttttgcactg actagtactg 7981 aataatacaa ccactcttat ttaatgttag tattatttat ttgacaactc agtgtctaac 8041 agcttgatat gcaggtcctt gcatcctaca tttctttagg aagttaccca tttgtaactt 8101 taaaaacagg aaaaatatca gttggcaaat gcaatctttt ttttttttaa gctaaaggtg 8161 ggtgaactgg aatgaaaatc tttctgatgt tgtgtctata agcagccttg atgggatatg 8221 ttagaagtgt catgaaagtg tgattctact tttgcagaaa aatctaaaga tcaatttata 8281 tagctttatt ttttacttta tcaaagtata cagaatttta atatgcatat attgtgtctg 8341 acttaaaatt ataatgtctg cgtcaccatt taaaatgtct gttcattatg taatgtaata 8401 aaagaaggtc ttcaaaaatg tatttaacat gaatggtatc catagttgtc atcatcataa 8461 atactggagt ttatttttaa attattaaac atagtaggtg cattaacata aatcagtctc 8521 cacacagtaa catttaactg ataattcatt aatcagcttt gaaaaattaa attgttaatt 8581 aaaccaatct aacatttcag taaagtttat tttgtatgct tctgttttta acttttattt 8641 ctgtagataa actgactgga taatattata ttggactttt ctctagatta tctaagcagg 8701 agacctgaat ctgcttgcaa taaagaataa aagtctgctt cagtttcttt ataaagaaac 8761 tcacacaa

In some embodiments of the methods of the disclosure, the wild type human ATP11A gene of the disclosure consists of or comprises the amino acid sequence (Genbank

Accession number: NP_115565.3, transcript variant 2):

(SEQ ID NO: 49)    1 mdcslvrtlv hrycageenw vdsrtiyvgh repppgaeay ipqrypdnri vsskytfwnf   61 ipknlfeqfr rvanfyflii flvqliidtp tspvtsglpl ffvitvtaik qgyedwlrhk  121 adnamnqcpv hfiqhgklvr kqsrklrvgd ivmvkedetf pcdliflssn rgdgtchvtt  181 asldgesshk thyavqdtkg fhteediggl hatieceqpq pdlykfvgri nvysdlndpv  241 vrplgsenll lrgatlknte kifgvaiytg metkmalnyq sksqkrsave ksmnaflivy  301 lciliskali ntvlkymwqs epfrdepwyn qkteserqrn lflkaftdfl afmvlfnyii  361 pvsmyvtvem qkflgsyfit wdedmfdeet gegplvntsd lneelgqvey iftdktgtlt  421 ennmefkecc ieghvyvphv icngqvlpes sgidmidssp svngrereel ffralclcht  481 vqvkdddsvd gprkspdggk scvyissspd evalvegvqr lgftylrlkd nymeilnren  541 hierfellei lsfdsvrrrm svivksatge iylfckgads sifprviegk vdqirarver  601 naveglrtlc vaykrliqee yegickllqa akvalqdrek klaeayeqie kdltllgata  661 vedrlqekaa dtiealqkag ikvwvltgdk metaaatcya cklfrrntql lelttkriee  721 qslhdvlfel sktvlrhsgs ltrdnlsgls admqdyglii dgaalslimk predgssgny  781 relfleicrs csavlccrma plqkaqivkl ikfskehpit laigdgandv smileahvgi  841 gvigkegrqa arnsdyaipk fkhlkkmllv hghfyyiris elvqyffykn vcfifpqfly  901 qffcgfsqqt lydtayltly nisftslpil lyslmeqhvg idvlkrdptl yrdvaknall  961 rwrvfiywtl lglfdalvff fgayfvfent tvtsngqifg nwtfgtlvft vmvftvtlkl 1021 aldthywtwi nhfviwgsll fyvvfsllwg gviwpflnyq rmyyvfiqml ssgpawlaiv 1081 llvtisllpd vlkkvlcrql wptatervqn gcaqprdrds eftplaslqs pgyqstcpsa 1141 awysshsqqv tlaawkekvs tepppilggs hhhcssipsh scprsrvgml v

In some embodiments of the methods of the disclosure, the wild type human IVD/DISP2 gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_002225.3, transcript variant 1):

(SEQ ID NO: 50)    1 tttccgcagt taggggctgc tatttcaacg cagggagata aaaagaaaaa aacacttgct   61 cttctacccc gctaaaaaca ctcatcctag ggagcacgcc agcatttgca gcgttcgggg  121 cagggccact cggcctgcgg ccgttgcact ggctggaagc tggcaggcga tcacggttga  181 ttggctcggg tgcggtccaa gggcagcaac gccttcggcg ggccgcctag ggtgattggc  241 tgctgcagcc caccccctag ccggtttggt gggcggcgaa gcctggattg gtggagctaa  301 gagctggctc agtttcagcg ctggctcttc gtgcatggca gagatggcga ctgcgactcg  361 gctgctgggg tggcgtgtgg cgagctggag gctgcggccg ccgcttgccg gcttcgtttc  421 ccagcgggcc cactcgcttt tgcccgtgga cgatgcaatc aatgggctaa gcgaggagca  481 gaggcagctt cgtcagacca tggctaagtt ccttcaggag cacctggccc ccaaggccca  541 ggagatcgat cgcagcaatg agttcaagaa cctgcgagaa ttttggaagc agctggggaa  601 cctgggcgta ttgggcatca cagcccctgt tcagtatggc ggctccggcc tgggctacct  661 ggagcatgtg ctggtgatgg aggagatatc ccgagcttcc ggagcagtgg ggctcagtta  721 cggtgcccac tccaacctct gcatcaacca gcttgtacgc aatgggaatg aggcccagaa  781 agagaagtat ctcccgaagc tgatcagtgg tgagtacatc ggagccctgg ccatgagtga  841 gcccaatgca ggctctgatg ttgtctctat gaagctcaaa gcggaaaaga aaggaaatca  901 ctacatcctg aatggcaaca agttctggat cactaatggc cctgatgctg acgtcctgat  961 tgtctatgcc aagacagatc tggctgctgt gccagcttct cggggcatca cagccttcat 1021 tgtggagaag ggtatgcctg gctttagcac ctctaagaag ctggacaagc tggggatgag 1081 gggctctaac acctgtgagc taatctttga agactgcaag attcctgctg ccaacatcct 1141 gggccatgag aataagggtg tctacgtgct gatgagtggg ctggacctgg agcggctggt 1201 gctggccggg gggcctcttg ggctcatgca agcggtcctg gaccacacca ttccctacct 1261 gcacgtgagg gaagcctttg gccagaagat cggccacttc cagttgatgc aggggaagat 1321 ggctgacatg tacacccgcc tcatggcgtg tcggcagtat gtctacaatg tcgccaaggc 1381 ctgcgatgag ggccattgca ctgctaagga ctgtgcaggt gtgattcttt actcagctga 1441 gtgtgccaca caggtagccc tggacggcat tcagtgtttt ggtggcaatg gctacatcaa 1501 tgactttccc atgggccgct ttcttcgaga tgccaagctg tatgagatag gggctgggac 1561 cagcgaggtg aggcggctgg tcatcggcag agccttcaat gcagactttc actagtcctg 1621 agacccttcg cccccttttc ctgcacctag tggcctttct tgggaagtag agatgtggcg 1681 gctttcccac cctgcccaca gcaggccctc ctgcccagct gctcttgtca gccctctggc 1741 ctctggatga ggttgagttc tccacaacag ctcccaagca tcatgggcct cgcagccggg 1801 cctgtgccac ggctagtgtt gtgtgattta aaatggactc agcaggaagc atattgtctg 1861 gggattgttg ggacaggttt tggtgactct gtgcccttgc tctctaactt ctgagcccac 1921 ctcccagggt aggcacctgg gggcatgcag gtgcccacct cccagggtag gcacctgggg 1981 gcatgcaggt acccacctct ttctcttggg tgaggctctg gcaaggagat ctctctgctc 2041 aagcacagca gaatcatggc ccctctccat gaattggaac ttggtacagg ttaagtatcc 2101 ctaatcctga aatctgaaac acttgtggtt ccaagcattt tggataaggc aaattcaact 2161 ttcagtctct tttctggggg aaaaaaataa taaacctagc ctagccaggc gtggtggctc 2221 atgcttgtaa tcccagcact tcaggaggct gagatgggtg gatcacctga ggtcaggagt 2281 tcaagaccag cctggccaac atgtggaaac ctcgcctcaa ctaaaaatag aaaaaaatta 2341 gttgggcatg gtggtgggca cctgtaatcc cagctacttc aggaggctga ggcaggagaa 2401 ttacttgaac ccaggaggcg gacgttgcag tgagccgagc ttgtgccatt gcactccagc 2461 ctgggcgaca agagcaaaac tcttcaaaaa acaaaacaaa acaaaaaaac cctggccctt 2521 gtttcttcca gtttctagag gtatcagctc ctagcagctt atgaacacat atgcttgctt 2581 ggccaggcaa ggtggtgtgt gcctgtaatc ccagcacttt gggaggccaa ggcaggtgga 2641 tcacttgcag tcaggagttc aagaccagcc tgtccaacgt ggtgaaaccc catctctact 2701 aaaaatacaa aaattagcca ggggtggtgg tgcacgtctg taatcccagc tactcaggag 2761 gctgaggcag gagaatcact tgaacccggg aggtggaggt tgcaatgagc caatatgaca 2821 ccgctgcagt ccagcctggg ccatagagtg agactctgtc tcaaaaaagg aaagaaaaat 2881 aggctgggca cagtgactca tgcctgtaat cccaacactt tgggaggccg aggcaggtgg 2941 atcacgaggt caggagttca agaccagcct ggccaagatg gtaaaacctc gtctctacta 3001 aaaatacaaa aattagccag gtgtggtggc aggctcctgt aatcccagct actcaggagg 3061 ctgaggcaga gaattgcttg aacccgggag gcagagtttg cagtgagcca agatcacacc 3121 actgcactcc agcttggacg acagagcgag actctgtctc aaaaaataat aggccaggca 3181 tggtggctca acgtctgtaa tcccagcact ttgggaggcc gaggcgggca gatcacaagg 3241 tcaggagttc gagaccagcc tgacgaccaa catggtgaaa cctcgtctct actaaaaata 3301 caaaaattag ccaggcctgg tggcacgcgc ctgtaatccc agttacacag aagactgagg 3361 caggagaatc gcttgaacgc aggaggcaga ggttgcagga gctgagatcg cgccattgca 3421 ctccagcctg ggcaacagag tgagactctg tctcaaaaaa taataataaa ataaatgaac 3481 acacatgctg ctgagtccgc agggggggca gagcagagga cagcgtgctt ttgtgtactg 3541 ttggaagact ggctcctcct gtacagcacc tctgagccct tgtgcaccgc cctgccacgg 3601 gcaccatcca gtcctggccg tgtgaccacc cacagctgac tgggcagcag gcacaggccc 3661 tacccgagca ggccggagtt ggctcgcatg actccagctg aggctgcctg tgtacatttc 3721 tccagatacc ctatggctaa ttttgttata actgcacagt ggctgctgcc attttgtatt 3781 aaatatattg tgaaacaaac ctatctgggg agaagcaatc tacttgccgc tgcttcctgt 3841 ctggatccag cttgtgtcct tggagagtgg ctggcccagg tcctattcct gtcctccagc 3901 ccgttctttc atgagggaca ggaaggtaaa atcagccctt aggagagagg tctcagcctc 3961 cctttcccag atctcccagt gagttttaaa ggaagcaggg agcccagagt gctaagttct 4021 tacagccaga aggaagctta tagatttctg aaaaccgccc ctttgttttt aaaaagatca 4081 acacaatttg actttctcaa ggtcaaaacg aactagaatc cagatctgct catggcaaaa 4141 atgggggtgt tctgagaatt ccagctttgg gccgcactgt acagcagtct ggatagagtg 4201 tgatctgaga agggaatggg tctgggttgt tccacccctt ccgagttcca aaaagaggga 4261 actggttttc ttggttctca gcccagcagc acctatcctg gctcttggtc ctggcctgca 4321 gccaagtgct gttcctagcc tgaggcttga gacaggtggg gttggctcct caccaacccc 4381 agttccgtcc catcctgagg gcaagatcct gggctcatag gcagtccctt tcacttcctt 4441 gtcttgctcc ctgctatgtt ggagatgaat gtgactaaaa gggccatctt gctggcttaa 4501 tgtgtggctg gagagaccag cctggagaca atgtggcaaa atggggcgct tcatccagtc 4561 tgtctaagcc ctgtcgactt ggggaggtga tttctttcct ggttctatat gtgaagcaaa 4621 ataaatgttt taaaattaaa agcaaaaaaa acaaaatgaa ccatgaaaaa aaa

In some embodiments of the methods of the disclosure, the wild type human IVD/DISP2 gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: transcript variant 1):

(SEQ ID NO: 51)   1 maematatrl lgwrvaswrl rpplagfvsq rahsllpvdd ainglseeqr qlrqtmakfl  61 qehlapkage idrsnefknl refwkqlgnl gvlgitapvq yggsglgyle hvlvmeeisr 121 asgavglsyg ahsnlcinql vrngneaqke kylpklisge yigalamsep nagsdvvsmk 181 lkaekkgnhy ilngnkfwit ngpdadvliv yaktdlaavp asrgitafiv ekgmpgfsts 241 kkldklgmrg sntcelifed ckipaanilg henkgvyvlm sgldlerlvl aggplglmqa 301 vldhtipylh vreafgqkig hfqlmqgkma dmytrlmacr qyvynvakac deghctakdc 361 agvilysaec atqvaldgiq cfggngyind fpmgrflrda klyeigagts evrrlvigra 421 fnadfh

In some embodiments of the methods of the disclosure, the wild type human IVD/DISP2 gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_001159508.1, transcript variant 2):

(SEQ ID NO: 27)    1 tttccgcagt taggggctgc tatttcaacg cagggagata aaaagaaaaa aacacttgct   61 cttctacccc gctaaaaaca ctcatcctag ggagcacgcc agcatttgca gcgttcgggg  121 cagggccact cggcctgcgg ccgttgcact ggctggaagc tggcaggcga tcacggttga  181 ttggctcggg tgcggtccaa gggcagcaac gccttcggcg ggccgcctag ggtgattggc  241 tgctgcagcc caccccctag ccggtttggt gggcggcgaa gcctggattg gtggagctaa  301 gagctggctc agtttcagcg ctggctcttc gtgcatggca gagatggcga ctgcgactcg  361 gctgctgggg tggcgtgtgg cgagctggag gctgcggccg ccgcttgccg gcttcgtttc  421 ccagcgggcc cactcgcttt tgcccgtgga cgatgcaatc aatgggctaa gcgaggagca  481 gaggcaggaa ttttggaagc agctggggaa cctgggcgta ttgggcatca cagcccctgt  541 tcagtatggc ggctccggcc tgggctacct ggagcatgtg ctggtgatgg aggagatatc  601 ccgagcttcc ggagcagtgg ggctcagtta cggtgcccac tccaacctct gcatcaacca  661 gcttgtacgc aatgggaatg aggcccagaa agagaagtat ctcccgaagc tgatcagtgg  721 tgagtacatc ggagccctgg ccatgagtga gcccaatgca ggctctgatg ttgtctctat  781 gaagctcaaa gcggaaaaga aaggaaatca ctacatcctg aatggcaaca agttctggat  841 cactaatggc cctgatgctg acgtcctgat tgtctatgcc aagacagatc tggctgctgt  901 gccagcttct cggggcatca cagccttcat tgtggagaag ggtatgcctg gctttagcac  961 ctctaagaag ctggacaagc tggggatgag gggctctaac acctgtgagc taatctttga 1021 agactgcaag attcctgctg ccaacatcct gggccatgag aataagggtg tctacgtgct 1081 gatgagtggg ctggacctgg agcggctggt gctggccggg gggcctcttg ggctcatgca 1141 agcggtcctg gaccacacca ttccctacct gcacgtgagg gaagcctttg gccagaagat 1201 cggccacttc cagttgatgc aggggaagat ggctgacatg tacacccgcc tcatggcgtg 1261 tcggcagtat gtctacaatg tcgccaaggc ctgcgatgag ggccattgca ctgctaagga 1321 ctgtgcaggt gtgattcttt actcagctga gtgtgccaca caggtagccc tggacggcat 1381 tcagtgtttt ggtggcaatg gctacatcaa tgactttccc atgggccgct ttcttcgaga 1441 tgccaagctg tatgagatag gggctgggac cagcgaggtg aggcggctgg tcatcggcag 1501 agccttcaat gcagactttc actagtcctg agacccttcg cccccttttc ctgcacctag 1561 tggcctttct tgggaagtag agatgtggcg gctttcccac cctgcccaca gcaggccctc 1621 ctgcccagct gctcttgtca gccctctggc ctctggatga ggttgagttc tccacaacag 1681 ctcccaagca tcatgggcct cgcagccggg cctgtgccac ggctagtgtt gtgtgattta 1741 aaatggactc agcaggaagc atattgtctg gggattgttg ggacaggttt tggtgactct 1801 gtgcccttgc tctctaactt ctgagcccac ctcccagggt aggcacctgg gggcatgcag 1861 gtgcccacct cccagggtag gcacctgggg gcatgcaggt acccacctct ttctcttggg 1921 tgaggctctg gcaaggagat ctctctgctc aagcacagca gaatcatggc ccctctccat 1981 gaattggaac ttggtacagg ttaagtatcc ctaatcctga aatctgaaac acttgtggtt 2041 ccaagcattt tggataaggc aaattcaact ttcagtctct tttctggggg aaaaaaataa 2101 taaacctagc ctagccaggc gtggtggctc atgcttgtaa tcccagcact tcaggaggct 2161 gagatgggtg gatcacctga ggtcaggagt tcaagaccag cctggccaac atgtggaaac 2221 ctcgcctcaa ctaaaaatag aaaaaaatta gttgggcatg gtggtgggca cctgtaatcc 2281 cagctacttc aggaggctga ggcaggagaa ttacttgaac ccaggaggcg gacgttgcag 2341 tgagccgagc ttgtgccatt gcactccagc ctgggcgaca agagcaaaac tcttcaaaaa 2401 acaaaacaaa acaaaaaaac cctggccctt gtttcttcca gtttctagag gtatcagctc 2461 ctagcagctt atgaacacat atgcttgctt ggccaggcaa ggtggtgtgt gcctgtaatc 2521 ccagcacttt gggaggccaa ggcaggtgga tcacttgcag tcaggagttc aagaccagcc 2581 tgtccaacgt ggtgaaaccc catctctact aaaaatacaa aaattagcca ggggtggtgg 2641 tgcacgtctg taatcccagc tactcaggag gctgaggcag gagaatcact tgaacccggg 2701 aggtggaggt tgcaatgagc caatatgaca ccgctgcagt ccagcctggg ccatagagtg 2761 agactctgtc tcaaaaaagg aaagaaaaat aggctgggca cagtgactca tgcctgtaat 2821 cccaacactt tgggaggccg aggcaggtgg atcacgaggt caggagttca agaccagcct 2881 ggccaagatg gtaaaacctc gtctctacta aaaatacaaa aattagccag gtgtggtggc 2941 aggctcctgt aatcccagct actcaggagg ctgaggcaga gaattgcttg aacccgggag 3001 gcagagtttg cagtgagcca agatcacacc actgcactcc agcttggacg acagagcgag 3061 actctgtctc aaaaaataat aggccaggca tggtggctca acgtctgtaa tcccagcact 3121 ttgggaggcc gaggcgggca gatcacaagg tcaggagttc gagaccagcc tgacgaccaa 3181 catggtgaaa cctcgtctct actaaaaata caaaaattag ccaggcctgg tggcacgcgc 3241 ctgtaatccc agttacacag aagactgagg caggagaatc gcttgaacgc aggaggcaga 3301 ggttgcagga gctgagatcg cgccattgca ctccagcctg ggcaacagag tgagactctg 3361 tctcaaaaaa taataataaa ataaatgaac acacatgctg ctgagtccgc agggggggca 3421 gagcagagga cagcgtgctt ttgtgtactg ttggaagact ggctcctcct gtacagcacc 3481 tctgagccct tgtgcaccgc cctgccacgg gcaccatcca gtcctggccg tgtgaccacc 3541 cacagctgac tgggcagcag gcacaggccc tacccgagca ggccggagtt ggctcgcatg 3601 actccagctg aggctgcctg tgtacatttc tccagatacc ctatggctaa ttttgttata 3661 actgcacagt ggctgctgcc attttgtatt aaatatattg tgaaacaaac ctatctgggg 3721 agaagcaatc tacttgccgc tgcttcctgt ctggatccag cttgtgtcct tggagagtgg 3781 ctggcccagg tcctattcct gtcctccagc ccgttctttc atgagggaca ggaaggtaaa 3841 atcagccctt aggagagagg tctcagcctc cctttcccag atctcccagt gagttttaaa 3901 ggaagcaggg agcccagagt gctaagttct tacagccaga aggaagctta tagatttctg 3961 aaaaccgccc ctttgttttt aaaaagatca acacaatttg actttctcaa ggtcaaaacg 4021 aactagaatc cagatctgct catggcaaaa atgggggtgt tctgagaatt ccagctttgg 4081 gccgcactgt acagcagtct ggatagagtg tgatctgaga agggaatggg tctgggttgt 4141 tccacccctt ccgagttcca aaaagaggga actggttttc ttggttctca gcccagcagc 4201 acctatcctg gctcttggtc ctggcctgca gccaagtgct gttcctagcc tgaggcttga 4261 gacaggtggg gttggctcct caccaacccc agttccgtcc catcctgagg gcaagatcct 4321 gggctcatag gcagtccctt tcacttcctt gtcttgctcc ctgctatgtt ggagatgaat 4381 gtgactaaaa gggccatctt gctggcttaa tgtgtggctg gagagaccag cctggagaca 4441 atgtggcaaa atggggcgct tcatccagtc tgtctaagcc ctgtcgactt ggggaggtga 4501 tttctttcct ggttctatat gtgaagcaaa ataaatgttt taaaattaaa agcaaaaaaa 4561 acaaaatgaa ccatg

In some embodiments of the methods of the disclosure, the wild type human IVD/DISP2 gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NP_001152980.1, transcript variant 2):

(SEQ ID NO: 28)   1 maematatrl lgwrvaswrl rpplagfvsq rahsllpvdd ainglseeqr qefwkqlgnl  61 gvlgitapvq yggsglgyle hvlvmeeisr asgavglsyg ahsnlcinql vrngneaqke 121 kylpklisge yigalamsep nagsdvvsmk lkaekkgnhy ilngnkfwit ngpdadvliv 181 yaktdlaavp asrgitafiv ekgmpgfsts kkldklgmrg sntcelifed ckipaanilg 241 henkgvyvlm sgldlerlvl aggplglmqa vldhtipylh vreafgqkig hfqlmqgkma 301 dmytrlmacr qyvynvakac deghctakdc agvilysaec atqvaldgiq cfggngyind 361 fpmgrflrda klyeigagts evrrlvigra fnadfh

In some embodiments of the methods of the disclosure, the wild type human DPP9 gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_139159.41:

(SEQ ID NO: 29)    1 caacttccgg gtcaaaggtg cctgagccgg cgggtcccct gtgtccgccg cggctgtcgt   61 cccccgctcc cgccacttcc ggggtcgcag tcccgggcat ggagccgcga ccgtgaggcg  121 ccgctggacc cgggacgacc tgcccagtcc ggccgccgcc ccacgtcccg gtctgtgtcc  181 cacgcctgca gctggaatgg aggctctctg gaccctttag aaggcacccc tgccctcctg  241 aggtcagctg agcggttaat gcggaaggtt aagaaactgc gcctggacaa ggagaacacc  301 ggaagttgga gaagcttctc gctgaattcc gagggggctg agaggatggc caccaccggg  361 accccaacgg ccgaccgagg cgacgcagcc gccacagatg acccggccgc ccgcttccag  421 gtgcagaagc actcgtggga cgggctccgg agcatcatcc acggcagccg caagtactcg  481 ggcctcattg tcaacaaggc gccccacgac ttccagtttg tgcagaagac ggatgagtct  541 gggccccact cccaccgcct ctactacctg ggaatgccat atggcagccg agagaactcc  601 ctcctctact ctgagattcc caagaaggtc cggaaagagg ctctgctgct cctgtcctgg  661 aagcagatgc tggatcattt ccaggccacg ccccaccatg gggtctactc tcgggaggag  721 gagctgctga gggagcggaa acgcctgggg gtcttcggca tcacctccta cgacttccac  781 agcgagagtg gcctcttcct cttccaggcc agcaacagcc tcttccactg ccgcgacggc  841 ggcaagaacg gcttcatggt gtcccctatg aaaccgctgg aaatcaagac ccagtgctca  901 gggccccgga tggaccccaa aatctgccct gccgaccctg ccttcttctc cttcatcaat  961 aacagcgacc tgtgggtggc caacatcgag acaggcgagg agcggcggct gaccttctgc 1021 caccaaggtt tatccaatgt cctggatgac cccaagtctg cgggtgtggc caccttcgtc 1081 atacaggaag agttcgaccg cttcactggg tactggtggt gccccacagc ctcctgggaa 1141 ggttcagagg gcctcaagac gctgcgaatc ctgtatgagg aagtcgatga gtccgaggtg 1201 gaggtcattc acgtcccctc tcctgcgcta gaagaaagga agacggactc gtatcggtac 1261 cccaggacag gcagcaagaa tcccaagatt gccttgaaac tggctgagtt ccagactgac 1321 agccagggca agatcgtctc gacccaggag aaggagctgg tgcagccctt cagctcgctg 1381 ttcccgaagg tggagtacat cgccagggcc gggtggaccc gggatggcaa atacgcctgg 1441 gccatgttcc tggaccggcc ccagcagtgg ctccagctcg tcctcctccc cccggccctg 1501 ttcatcccga gcacagagaa tgaggagcag cggctagcct ctgccagagc tgtccccagg 1561 aatgtccagc cgtatgtggt gtacgaggag gtcaccaacg tctggatcaa tgttcatgac 1621 atcttctatc ccttccccca atcagaggga gaggacgagc tctgctttct ccgcgccaat 1681 gaatgcaaga ccggcttctg ccatttgtac aaagtcaccg ccgttttaaa atcccagggc 1741 tacgattgga gtgagccctt cagccccggg gaagatgaat ttaagtgccc cattaaggaa 1801 gagattgctc tgaccagcgg tgaatgggag gttttggcga ggcacggctc caagatctgg 1861 gtcaatgagg agaccaagct ggtgtacttc cagggcacca aggacacgcc gctggagcac 1921 cacctctacg tggtcagcta tgaggcggcc ggcgagatcg tacgcctcac cacgcccggc 1981 ttctcccata gctgctccat gagccagaac ttcgacatgt tcgtcagcca ctacagcagc 2041 gtgagcacgc cgccctgcgt gcacgtctac aagctgagcg gccccgacga cgaccccctg 2101 cacaagcagc cccgcttctg ggctagcatg atggaggcag ccagctgccc cccggattat 2161 gttcctccag agatcttcca tttccacacg cgctcggatg tgcggctcta cggcatgatc 2221 tacaagcccc acgccttgca gccagggaag aagcacccca ccgtcctctt tgtatatgga 2281 ggcccccagg tgcagctggt gaataactcc ttcaaaggca tcaagtactt gcggctcaac 2341 acactggcct ccctgggcta cgccgtggtt gtgattgacg gcaggggctc ctgtcagcga 2401 gggcttcggt tcgaaggggc cctgaaaaac caaatgggcc aggtggagat cgaggaccag 2461 gtggagggcc tgcagttcgt ggccgagaag tatggcttca tcgacctgag ccgagttgcc 2521 atccatggct ggtcctacgg gggcttcctc tcgctcatgg ggctaatcca caagccccag 2581 gtgttcaagg tggccatcgc gggtgccccg gtcaccgtct ggatggccta cgacacaggg 2641 tacactgagc gctacatgga cgtccctgag aacaaccagc acggctatga ggcgggttcc 2701 gtggccctgc acgtggagaa gctgcccaat gagcccaacc gcttgcttat cctccacggc 2761 ttcctggacg aaaacgtgca ctttttccac acaaacttcc tcgtctccca actgatccga 2821 gcagggaaac cttaccagct ccagatctac cccaacgaga gacacagtat tcgctgcccc 2881 gagtcgggcg agcactatga agtcacgttg ctgcactttc tacaggaata cctctgagcc 2941 tgcccaccgg gagccgccac atcacagcac aagtggctgc agcctccgcg gggaaccagg 3001 cgggagggac tgagtggccc gcgggcccca gtgaggcact ttgtcccgcc cagcgctggc 3061 cagccccgag gagccgctgc cttcaccgcc ccgacgcctt ttatcctttt ttaaacgctc 3121 ttgggtttta tgtccgctgc ttcttggttg ccgagacaga gagatggtgg tctcgggcca 3181 gcccctcctc tccccgcctt ctgggaggag gaggtcacac gctgatgggc actggagagg 3241 ccagaagaga ctcagaggag cgggctgcct tccgcctggg gctccctgtg acctctcagt 3301 cccctggccc ggccagccac cgtccccagc acccaagcat gcaattgcct gtcccccccg 3361 gccagcctcc ccaacttgat gtttgtgttt tgtttggggg gatatttttc ataattattt 3421 aaaagacagg ccgggcgcgg tggctcacgt ctgtaatccc agcactttgg gaggctgagg 3481 cgggcggatc acctgaggtt gggagttcaa gaccagcctg gccaacatgg ggaaaccccg 3541 tctctactaa aaatacaaaa aattagccgg gtgtggtggc gcgtgcctat aatcccagct 3601 actcgggagg ctgaggcagg agaatcgctt gaacccggga ggtggaggtt gcggtgagcc 3661 aagatcgcac cattgcactc cagcctgggc aacaagagcg aaactctgtc tcaaaataaa 3721 taaaaaataa aagacagaaa gcaaggggtg cctaaatcta gacttggggt ccacaccggg 3781 cagcggggtt gcaacccagc acctggtagg ctccatttct tcccaagccc gagcagaggg 3841 tcatgcgggc cccacaggag aagcggccag ggcccgcggg gggcaccacc tgtggacagc 3901 cctcctgtcc ccaagctttc aggcaggcac tgaaacgcac cgaacttcca cgctctgctg 3961 gtcagtggcg gctgtcccct ccccagccca gccgcccagc cacatgtgtc tgcctgaccc 4021 gtacacacca ggggttccgg ggttgggagc tgaaccatcc ccacctcagg gttatatttc 4081 cctctcccct tccctccccg ccaagagctc tgccaggggc gggcaaaaaa aaaagtaaaa 4141 agaaaagaaa aaaaaaaaaa agaaacaaac cacctctaca tattatggaa agaaaatatt 4201 tttgtcgatt cttattcttt tataattatg cgtggaagaa gtagacacat taaacgattc 4261 cagttggaaa aaaaaaaaaa aaaaaa

In some embodiments of the methods of the disclosure, the wild type human

DPP9 gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_631898.3):

(SEQ ID NO: 30)   1 mrkvkklrld kentgswrsf slnsegaerm attgtptadr gdaaatddpa arfqvqkhsw  61 dglrsiihgs rkysglivnk aphdfqfvqk tdesgphshr lyylgmpygs rensllysei 121 pkkvrkeall llswkqmldh fqatphhgvy sreeellrer krlgvfgits ydfhsesglf 181 lfqasnslfh crdggkngfm vspmkpleik tqcsgprmdp kicpadpaff sfinnsdlwv 241 anietgeerr ltfchqglsn vlddpksagv atfviqeefd rftgywwcpt aswegseglk 301 tlrilyeevd esevevihvp spaleerktd syryprtgsk npkialklae fqtdsqgkiv 361 stqekelvqp fsslfpkvey iaragwtrdg kyawamfldr pqqwlqlvll ppalfipste 421 neeqrlasar avprnvqpyv vyeevtnvwi nvhdifypfp qsegedelcf lranecktgf 481 chlykvtavl ksqgydwsep fspgedefkc pikeeialts gewevlarhg skiwvneetk 541 lvyfqgtkdt plehhlyvvs yeaageivrl ttpgfshscs msqnfdmfvs hyssystppc 601 vhvyklsgpd ddplhkqprf wasmmeaasc ppdyvppeif hfhtrsdvrl ygmiykphal 661 qpgkkhptvl fvyggpqvql vnnsfkgiky lrlntlaslg yavvvidgrg scqrglrfeg 721 alknqmgqve iedqveglqf vaekygfidl srvaihgwsy ggflslmgli hkpqvfkvai 781 agapvtvwma ydtgyterym dvpennqhgy eagsvalhve klpnepnrll ilhgfldenv 841 hffhtnflvs qliragkpyq lqiypnerhs ircpesgehy evtllhflqe yl

In some embodiments of the methods of the disclosure, the wild type human SIGLEC14 gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_001098612.1):

(SEQ ID NO: 31)    1 actcaccctc cggcttcctg tcggggcttt ctcagcccca ccccacgttt ggacatttgg   61 agcatttcct tccctgacag ccggacctgg gactgggctg gggccctggc ggatggagac  121 atgctgcccc tgctgctgct gcccctgctg tggggggggt ccctgcagga gaagccagtg  181 tacgagctgc aagtgcagaa gtcggtgacg gtgcaggagg gcctgtgcgt ccttgtgccc  241 tgctccttct cttacccctg gagatcctgg tattcctctc ccccactcta cgtctactgg  301 ttccgggacg gggagatccc atactacgct gaggttgtgg ccacaaacaa cccagacaga  361 agagtgaagc cagagaccca gggccgattc cgcctccttg gggatgtcca gaagaagaac  421 tgctccctga gcatcggaga tgccagaatg gaggacacgg gaagctattt cttccgcgtg  481 gagagaggaa gggatgtaaa atatagctac caacagaata agctgaactt ggaggtgaca  541 gccctgatag agaaacccga catccacttt ctggagcctc tggagtccgg ccgccccaca  601 aggctgagct gcagccttcc aggatcctgt gaagcgggac cacctctcac attctcctgg  661 acggggaatg ccctcagccc cctggacccc gagaccaccc gctcctcgga gctcaccctc  721 acccccaggc ccgaggacca tggcaccaac ctcacctgtc aggtgaaacg ccaaggagct  781 caggtgacca cggagagaac tgtccagctc aatgtctcct atgctccaca gaacctcgcc  841 atcagcatct tcttcagaaa tggcacaggc acagccctgc ggatcctgag caatggcatg  901 tcggtgccca tccaggaggg ccagtccctg ttcctcgcct gcacagttga cagcaacccc  961 cctgcctcac tgagctggtt ccgggaggga aaagccctca atccttccca gacctcaatg 1021 tctgggaccc tggagctgcc taacatagga gctagagagg gaggggaatt cacctgccgg 1081 gttcagcatc cgctgggctc ccagcacctg tccttcatcc tttctgtgca gagaagctcc 1141 tcttcctgca tatgtgtaac tgagaaacag cagggctcct ggcccctcgt cctcaccctg 1201 atcagggggg ctctcatggg ggctggcttc ctcctcacct atggcctcac ctggatctac 1261 tataccaggt gtggaggccc ccagcagagc agggctgaga ggcctggctg agcccctccc 1321 gctcaagaca gaactgaggt gtggacactt agccctgtgg gacacatgca ggacatcact 1381 gtcagcttct ttctggaagc tcacatccca ctgactaccc ctcttttcct tcctgcccca 1441 taccccttct acttattccc ctctgcttgt gagtcttgcc ccaccacacc tgcatcccca 1501 tctgcacccc atcccctctc cacctgccct tctcttccct ctccatccac catctccagc 1561 cctgtgaagg gaatgtactt tcggtcttat acccccatta cccattaccc aaaagttacc 1621 tttttttttt tttttttttt ttgagacaga gtctcactct gttgcacagg ctggagttca 1681 gtggcacaat ctccgttcac tgcaacctcc acctctgggg ttcaagcaat tctcctgcct 1741 cagcctccct agtagctggg attacaggtg cctgccacca catccagtta attttttttt 1801 tttgtatgtt agtagagatg gggttttacc atgttggcca ggtctcgaac tcctgacctc 1861 aagcaatcca ctgcattggc ctcccaaagt gctggcatta caggtatgag ccaccgtgcc 1921 tggctgccaa aagttacctt cttaacactt gaatttctgg tctcctcagc ttccctatcc 1981 atataggcac agagaggcag catttgtttt ccagttaaaa ctctacctca ttgtgattat 2041 tatccaatac aattgttaca aaataagtaa aacttttatg aaacaataca acataactga 2101 ttttactctt taa

In some embodiments of the methods of the disclosure, the wild type human SIGLEC14 gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_001092082.1):

(SEQ ID NO: 32)   1 mlpllllpll wggslqekpv yelqvqksvt vqeglcvlvp csfsypwrsw ysspplyvyw  61 frdgeipyya evvatnnpdr rvkpetqgrf rllgdvqkkn cslsigdarm edtgsyffry 121 ergrdvkysy qqnklnlevt aliekpdihf leplesgrpt rlscslpgsc eagppltfsw 181 tgnalspldp ettrsseltl tprpedhgtn ltcqvkrqga qvttertvql nvsyapqnla 241 isiffrngtg talrilsngm svpiqegqsl flactvdsnp paslswfreg kalnpsqtsm 301 sgtlelpnig areggeftcr vqhplgsqhl sfilsvqrss sscicvtekq qgswplvltl 361 irgalmgagf lltygltwiy ytrcggpqqs raerpg

In some embodiments of the methods of the disclosure, the wild type human ADM2 gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_001253845.1):

(SEQ ID NO: 33)    1 cgcccacgcc cggcgccccg accgcggagg actccccgag ccccgcccgc catggcccgg   61 atcccgacgg ccgccctggg ttgcatcagc ctcctctgcc tgcagctccc tggctcgctg  121 tcccgcagcc tgggcgggga cccgcgaccc gtcaaaccca gggagccccc agcccggagc  181 ccttccagca gcctgcagcc caggcacccc gcaccccgac ctgtggtctg gaagcttcac  241 cgggccctcc aggcacagag gggtgccggc ctggcccctg ttatgggtca gcctctccgg  301 gatggtggcc gccaacactc gggcccccga agacactcgg gcccccgcag gacccaagcc  361 cagctcctgc gagtgggctg tgtgctgggc acctgccagg tgcagaatct cagccaccgc  421 ctgtggcaac tcatgggacc ggccggccgg caggactcag ctcctgtgga ccccagcagc  481 ccccacagct atggctgagg tggggccggg ccacacccct gcccatccca gccagggtgc  541 tgtgcccccg tccagagctg cagctgagcc ccatctgaag cccagtccct cggagctgca  601 gacagcaggt cctgcagcaa caatacctgc acggctttgc acacgtaaac ctaggctggt  661 ctacacgcag tgctggtacg tcaaggagcc taaacaccct gaaattgtga ccccctgggg  721 gacagctgcc agacacagct ggcggcagca ccagatgcta agcgcttcag agaggaggtg  781 tctgcccaga gatgtggagc agaagctggg ccctgaacac acggggccat gtctggacga  841 gcaggggaga gaggctgaac tggccagaag tggcccctcc gctgctggtc cagtcagact  901 gaagcccggc cttgtgcctg ggctgttcct gctctcatgc acaaccagcc cttccacgtg  961 cctgcctgtg ggacaggagg gggagcgtgg gatgctgtag cccccggggt tgggcaaggg 1021 aaggatggtg gccctccaga ggtcatgaag ggacctctgt ggctccagct gccaaccctg 1081 gagcccagac cgaggtggcc atggagactc cacctggatc ccctgtagga ggccagggag 1141 gggaactcag cagttcagga gccaccccaa accattctgg gacagggaca cccctttcta 1201 ccccagggca gggcagggct gggtggggca agatccccca gcccgactag acccacctca 1261 cctgaagggg gtgagaccct tgttggcagc cagacaaggg tggggctcca caggcagcac 1321 aggcgcccca ccaccaccca gtttggggac ccagtgggac caggtgcggg ggcagagggt 1381 gacttaccaa gagccaggga gggcagccca ggcccaagtg acagcaagaa caagaaccac 1441 tgccggcgtg cacagacttg gtgtgtgtcc ttccctgggg ggacggggga ctcacatgtg 1501 cctgccactg gagcctctca accgtccagc agaacacggg gttcagaaag ggctccttct 1561 gctatttagc gaacactgag catttaattt acaaatgttt gctagggtca ccctctcggc 1621 catcccacga gggtcgccat gatcacccca actctagagg ccgcagcaga gctcaggaca 1681 ttcccccaca gagcttgccc ctcagttcct acctccaagg gggagggtcc tggaagcgcc 1741 cacccaggcg ccgcccctgt gcttgctccc cgagctcagg gattgccgag tccacgtaac 1801 tgacctgtac tccacgaggc cctgtgggaa cggtccaggc tggtcctgcc ctgtggaggc 1861 ctccgtgcac tgagagatgt actaggattg cagcaaaggt ggtcagggtg atgggccgca 1921 cagcgaggca gtcaaggcca gctccctggg agaagcactg ggtcaggtga ggtctgagga 1981 cagcaggcct tccctagggg aaggagctgg gagtgccaag gccccaggtg cacaggaggc 2041 gtggctgctg agaggctgca gggtggaggg gcctcggcct cagagtcatg tgccctgtga 2101 ccactgaagg gtgtcagcag agcacacggc atgaggacag agggaggggc acggggagtg 2161 aaggaggggg ccctggggca aggctcgggg gtcaggagct cagcgtccgc tactcagccc 2221 agccaaaacc ctcccagacg tctcctctcc tgcctgggca aagtccagct tggcaccccg 2281 tctggggcct gcctgtggtc agggccaagt gttccctcct ccaggaaagc ctttaccctc 2341 ctcatgccct gtagtcagga ggccgcctgc tgtaaccctc cgtgtcgcct cgggtgcgaa 2401 atcagaccca cctgacacca tcacgcggag gcccagcagc acctgcaccc acttccagct 2461 gctctggcca aaatctccgc tcggccaggc cccgtggctc acacctgtaa tcctagcaca 2521 ttgggaggcc aaggcaggca catcacctga gttcaggagt tcaagaccag cctggccaac 2581 atggtgaaat cccgtctcta ctaaaaacag aaaattatcc gggcgtggtg gcacatgact 2641 gtaatcccag ctactcagga ggctgaggca ggaggatcac ttgaacctgg gaggcggagg 2701 ttgcagtgag ctgagattgc gccattgcac tccagcctgg gcaacaagag caaaattctg 2761 cctcaaaaaa aaaaatagta ataatacaaa aattagctgg gcgtggtggc acatgccagt 2821 aattccatct actcgggagg ctgaggcagg agaatcgtct aagcccggga ggtggaggtt 2881 gcagtgagcc cagatggcgc tgctgcactc aagcttggat gacagagcaa gactccgttt 2941 caaaaaaaaa aaacctcctc tcttccttca caccttcctc tgaatcccac ccggtcccac 3001 ctcctgaacc tatccagaca ccttctcctg acccaggcac cacctgcttt cggggcgatg 3061 gccgtagcct cctcccaggc acctgtctgc atccctctgg ccagtgcatg ctgagcacgt 3121 gacctacccg tgttgggaca cgtgaggata cagccttgac ccccaggggc tgacattcta 3181 gggggagata gaaggagaca aacgtagaag gtagaataag tgggtggtgg agtggcaggg 3241 agtgctgagt gccacaggaa gtcagacaag gaaggagagt gtggggcagg tgccgtttaa 3301 atggggggcg ctggggtctc ctcacagttg cttctcagct cagctgtgcc aggatcttgt 3361 tgagtcaggt cagctgccca cagccctctt gcctgacccc tgaagcccag aactctgatc 3421 ttcacagccc taggtatggc cccagcaccc cactgccctc tctcctgccc cagccgactg 3481 ctgttcccag acttccctgg ccacgctcca agacgccagc tctgccgcgg gcactttgtt 3541 ctcacggtgt cctccatgcc tgcagggccc atgcatggga agttgcgttg gcggcctggg 3601 tgttggcggt tccgtgcctg ctccaactct ccgtgaggcc cctctcccag agcctgacac 3661 actctgtggc cgaactctag gcaggtgccc ctgagtcctt tcctcgacga ggcctgaccc 3721 catccccatc ctcgctgggc ccgccgaccc cggtgttagc aagaatcctc taaatcagtt 3781 tatggagaat tacccaccct cgatatctga tcccattcct catctcccac ccttgatctc 3841 atcaccctgc cggcctcctg caagatcctc attgagccac tccagtgaga atccccctac 3901 cctcgaaggc cgccctaaca acttcccatc cgctgacccc tccaacgcca tcaatctcca 3961 gctgtggttg ttgaactcgg aggtgagctc ctctcaccac tctcttgaat aaagcttttc 4021 tcaccatttt aaaaaaaaaa aaaaa

In some embodiments of the methods of the disclosure, the wild type human ADM2 gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_001240774.1):

(SEQ ID NO: 34)   1 mariptaalg cisllclqlp gslsrslggd prpvkprepp arspssslqp rhpaprpvvw  61 klhralqaqr gaglapvmgq plrdggrqhs gprrhsgprr tqaqllrvgc vlgtcqvqnl 121 shrlwqlmgp agrqdsapvd pssphsyg

In some embodiments of the methods of the disclosure, the wild type human TSPAN5 gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_005723.3):

(SEQ ID NO: 35)    1 aggcgggcgg agcgaggggt gggagggcgc gcgcgaacgg gcgggcgagc aagcgagcgg   61 cgtctccacc agcatctgcc gcggccgcct ttgcccgaag cccggggacg aaccgacgga  121 ccgaccgcct ggcgcacgga cgcgggcgct cgctttgtgt tcggggctag cgtcggcgag  181 gcttgagctt gcagcgcgcg gcttccctgc tttctcgcgg ccaccccggc tccggcggcc  241 tcggcgcgcg aggggctgga ggtgcgggag ccgctctccg ccggtcggtc cccgcgcggc  301 tgagcccagg ccgccagcgc cgcggccccg tgcggtgtcc ctgagctcct gctccccgcc  361 gggctgctcc gagcaacggt gcttcggagc tccaaactcg ggctgccggg gcaagtgtct  421 tcatgaaccc agaggatgtc cgggaagcac tacaagggtc ctgaagtcag ttgttgcatc  481 aaatacttca tatttggctt caatgtcata ttttggtttt tgggaataac atttcttgga  541 attggactgt gggcatggaa tgaaaaagga gttctgtcca acatctcttc catcaccgat  601 ctcggcggct ttgacccagt ttggctcttc cttgtggtgg gaggagtgat gttcattttg  661 ggatttgcag ggtgcattgg agcgctacgg gaaaacactt tccttctcaa gtttttttct  721 gtgttcctgg gaattatttt cttcctggag ctcactgccg gagttctagc atttgttttc  781 aaagactgga tcaaagacca gctgtatttc tttataaaca acaacatcag agcatatcgg  841 gatgacattg atttgcaaaa cctcatagac ttcacccagg aatattggca gtgctgtggg  901 gcttttggag ctgatgattg gaacctaaat atttacttca attgcacaga ttccaatgca  961 agtcgagagc gatgtggcgt tccattctcc tgctgcacta aagatcccgc agaagatgtc 1021 atcaacactc agtgtggcta tgatgccagg caaaaaccag aagttgacca gcagattgta 1081 atctacacga aaggctgtgt gccccagttt gagaagtggt tgcaggacaa tttaaccatc 1141 gttgctggta ttttcatagg cattgcattg ctgcagatat ttgggatatg cctggcccag 1201 aatttggtta gcgatatcga agctgtcagg gcgagctggt agaccccctg caaccgctgc 1261 tgcaagacac tggacagacc cagctttcgg gaccctcccg cgtgccgaac tgatcttcga 1321 gctgcatgga cctaatcaca gatgcagcct gcagtctcgc ctaatggagc tgccattagg 1381 ggagtgtaaa actgggaaat gctgctcact gacagaatta aaaaaaaaaa taaccagtat 1441 gaaagtcgtt gcgccgtgaa tctctactgt agccatgaat ttatggacag ttagatgctt 1501 accaaaaaag aaaaaaaggg agggtagggg acccagatgt acttgaatgt gcagaaaata 1561 cattcttgtc ctcatcttcc gtaattggag ggctgggaga ggcagctttg ctcttcacca 1621 caccttggac ggaccacctt ctttctgttc catggcctga aggagtgcat ctcctcaaag 1681 actcagcccc tcacctggga gggcagtggt ttgtgggcat ccctccatgt acattttagg 1741 aaacacttgc aactctcatc tgaagaagaa aacaactcat ctttgggttc agattttgtg 1801 atggtattca gcaagtcact tgggcgagca cacttggtct atcctggaaa gtctccttat 1861 aagagaagtt gtgtatttca tgtgcaccga gcaagggcat tggaagacgt catgaggctg 1921 tattttagca ggactgatcg tttttctaag tagacctgag ctttgtttat cagtgaaatt 1981 caaggagaaa atgaggttaa tgaagaggta tcagttaaat atccccttct tctcaccctg 2041 ccaaaattag cagttggatt tttggaaact ctggaatatt ctgggtcatt ttgttttgta 2101 tgtttgttgt ttttcgtctt ccaaaggtga aagctatgat acagttccac ttaaatttta 2161 gtgttttctt actcagctca agcattaatt tttgattaag tcttaatctg catgacctgt 2221 gaatctgaat ccatcatctc cctttcctgc cagcttttct acaaacattg aaatatgtta 2281 tttggtcagc acttatttcc taggttcaca gccttgggag gttgtggcat gtcctcccag 2341 tctggctggg aagagaccag ctgtaccatc caaatgcttc cctggtcttg atgatctctt 2401 ccagagtcga tctgagtggc cttttctgca ccctcccctt ctttctcttt gaatggaatt 2461 aaacccaatt tggaaacaac attgacccag tcaaaagctt ctaatggttt ctttttcttc 2521 ctccagtttt agtttgcttt tattaaaaaa agaaaatagt gcatggccat agctccttca 2581 gttctcttat tgcagactaa ccatcaggat ggtatcaaag cacaaatact ttggagggga 2641 atgcgttgaa ctggggcaag tactctgtaa cacaaagtgg gaaaccactt cctggtgctg 2701 ccgctcctgc ccccacttta ggtgggaggg acgagttttg ccctctagat tttaatccag 2761 ctggtgtcca ccggatgttg ccctcctggg gagcagatat cagtctgtgg aactctggga 2821 aaaccacagg cacatttttc ggtgcggaca gatttgccag cacataactg ggcagccagc 2881 tagaatactt tgtggaaatt aagcgaggtt ttccatttca gccccatggt gcatggtggt 2941 ggccgatgaa tgtgtcagtc tgctcagaga aaggacaaaa aggaaattat tttcaaaact 3001 gtgttcactg tttgggtgtg tgtatggctc tgcatgtgtg tgtttttgtc tctgtatagg 3061 tagaggtatt cacatcttac tccgactgta aggttgtctt acttcatctc tgcccccacc 3121 acagttgcca ttttgtaatg tccttccaac atggagaaga cacgagctct ctccagttgg 3181 catcatttgt cttttttgtt gattgcctca ttctccagtg aactccatct ggccaattga 3241 ttcagaatca ggcaagatcc ctgccctttg gcacatccac tgaaaggcca aacagcaagt 3301 ccgagtgagt tttaaatatt aattaatcac cctttatttt ttacacttga gagtgattgt 3361 aataaaggct gtcattaata aacttggttc taccttaaaa aaaaaa

In some embodiments of the methods of the disclosure, the wild type human TSPAN5 gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_005714.2):

(SEQ ID NO: 52)   1 msgkhykgpe vsccikyfif gfnvifwflg itflgiglwa wnekgvlsni ssitdlggfd  61 pvwlflvvgg vmfilgfagc igalrentfl lkffsvflgi iffleltagv lafvfkdwik 121 dqlyffinnn irayrddidl qnlidftqey wqccgafgad dwnlniyfnc tdsnasrerc 181 gvpfscctkd paedvintqc gydarqkpev dqqiviytkg cvpqfekwlq dnitivagif 241 igiallqifg iclaqnlvsd ieavrasw

In some embodiments of the methods of the disclosure, the wild type human CAMKK1 gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_032294.2, transcript variant 1):

(SEQ ID NO: 53)    1 ctgggcccca gcgaggcggt ggggcggggc ggggcggggc ggggcgcgca gcaggagcga   61 gtggggccgc ccgccgggcc gcggacactg tcgcccggcg cccaggttcc caacaaggct  121 acgcagaaga acccccttga ctgaagcaat ggaggggggt ccagctgtct gctgccagga  181 tcctcgggca gagctggtag aacgggtggc agccatcgat gtgactcact tggaggaggc  241 agatggtggc ccagagccta ctagaaacgg tgtggacccc ccaccacggg ccagagctgc  301 ctctgtgatc cctggcagta cttcaagact gctcccagcc cggcctagcc tctcagccag  361 gaagctttcc ctacaggagc ggccagcagg aagctatctg gaggcgcagg ctgggcctta  421 tgccacgggg cctgccagcc acatctcccc ccgggcctgg cggaggccca ccatcgagtc  481 ccaccacgtg gccatctcag atgcagagga ctgcgtgcag ctgaaccagt acaagctgca  541 gagtgagatt ggcaagggtg cctacggtgt ggtgaggctg gcctacaacg aaagtgaaga  601 cagacactat gcaatgaaag tcctttccaa aaagaagtta ctgaagcagt atggctttcc  661 acgtcgccct cccccgagag ggtcccaggc tgcccaggga ggaccagcca agcagctgct  721 gcccctggag cgggtgtacc aggagattgc catcctgaag aagctggacc acgtgaatgt  781 ggtcaaactg atcgaggtcc tggatgaccc agctgaggac aacctctatt tggtgtttga  841 cctcctgaga aaggggcccg tcatggaagt gccctgtgac aagcccttct cggaggagca  901 agctcgcctc tacctgcggg acgtcatcct gggcctcgag tacttgcact gccagaagat  961 cgtccacagg gacatcaagc catccaacct gctcctgggg gatgatgggc acgtgaagat 1021 cgccgacttt ggcgtcagca accagtttga ggggaacgac gctcagctgt ccagcacggc 1081 gggaacccca gcattcatgg cccccgaggc catttctgat tccggccaga gcttcagtgg 1141 gaaggccttg gatgtatggg ccactggcgt cacgttgtac tgctttgtct atgggaagtg 1201 cccattcatc gacgatttca tcctggccct ccacaggaag atcaagaatg agcccgtggt 1261 gtttcctgag gagccagaaa tcagcgagga gctcaaggac ctgatcctga agatgttaga 1321 caagaatccc gagacgagaa ttggggtgcc agacatcaag ttgcaccctt gggtgaccaa 1381 gaacggggag gagccccttc cttcggagga ggagcactgc agcgtggtgg aggtgacaga 1441 ggaggaggtt aagaactcag tcaggctcat ccccagctgg accacggtga tcctggtgaa 1501 gtccatgctg aggaagcgtt cctttgggaa cccgtttgag ccccaagcac ggagggaaga 1561 gcgatccatg tctgctccag gaaacctact ggtgaaagaa gggtttggtg aagggggcaa 1621 gagcccagag ctccccggcg tccaggaaga cgaggctgca tcctgagccc ctgcatgcac 1681 ccagggccac ccggcagcac actcatcccg cgcctccaga ggcccacccc tcatgcaaca 1741 gccgcccccg caggcagggg gctggggact gcagccccac tcccgcccct cccccatcgt 1801 gctgcatgac ctccacgcac gcacgtccag ggacagactg gaatgtatgt catttggggt 1861 cttgggggca gggctcccac gaggccatcc tcctcttctt ggacctcctt ggcctgaccc 1921 attctgtggg gaaaccgggt gcccatggag cctcagaaat gccacccggc tggttggcat 1981 ggcctggggc aggaggcaga ggcaggagac caagatggca ggtggaggcc aggcttacca 2041 caacggaaga gacctcccgc tggggccggg caggcctggc tcagctgcca caggcatatg 2101 gtggagaggg gggtaccctg cccaccttgg ggtggtggca ccagagctct tgtctattca 2161 gacgctggta tgggggctcg gacccctcac tggggacagg gccagtgttg gagaattctg 2221 attccttttt tgttgtcttt tacttttgtt tttaacctgg gggttcgggg agaggccctg 2281 cttgggaaca tctcacgagc tttcctacat cttccgtggt tcccagcaca gcccaagatt 2341 atttggcagc caagtggatg gaactaactt tcctggactg tgtttcgcat tcggcgttat 2401 ctggaaagtg gactgaacgg aatcaagctc tgagcagagg cctgaagcgg aagcaccaca 2461 tcgtccctgc ccatctcact ctctcccttg atgatgcccc tagagctgag gctggagaag 2521 acaccagggc tgactttgac cgagggccat ggacgcgaca ggcctgtggc cctgcgcatg 2581 ctgaaataac tggaacccag cctctcctcc tacaccggcc tacccatctg ggcccaagag 2641 ctgcactcac actcctacaa cgaaggacaa actgtccagg tcggagggat cacgagacac 2701 agaacctgga ggggtgtgca cgctggcagg tggcctctgc ggcaattgcc tcaccctgag 2761 gacatcagca gtcagcctgc tcagagcggg ggtgctggag cgcgtgcaga cacagctctt 2821 ccggagcagc cttcaccttc tctctgggat cagtgtccgg ctggccgacg tggcatttgc 2881 tgaccgaatg ctcatagagg ttgaccccca cagggtcacg caggactcgg acactgccct 2941 ggaaacatgg atggacaagg gcttttggcc acaggtgtgg gtgtcctgtt ggaggagggc 3001 ttgtttggag aagggaggct ggctggggga gaaacccgga tcccgctgca tctccgcgcc 3061 tgtgggtgca tgtcgcgtgc tcatctgttg cacacagctc actcgtatgt cctgcactgg 3121 tacatgcatc tgtaatacag tttctacgtc tatttaaggc taggagccga atgtgcccca 3181 ttgtcagtgg gtccacgttt ctccccggct cctctgggct aaggcagtgt ggcccgaagc 3241 ttaaaaagtt actcggtact gtttttaaga acacttttat agagttagtg gaaggcaagt 3301 taagagccaa tcactgatcc ccaagtgttt cttgagcatc tggtctgggg ggaccacttt 3361 gatcggaccc acccttggaa agctcagggg taggcccagg tgggatgctc accctgtcac 3421 tgagggtttt ggttggcatc gttgtttttg aatgtagcac aagcgatgag caaactctat 3481 aagagtgttt taaaaattaa cttcccagga agtgagttaa aaacaataaa agccctttct 3541 tgagttaaaa agaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaa

In some embodiments of the methods of the disclosure, the wild type human CAMKK1 gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_115670.1, transcript variant 1):

(SEQ ID NO: 54)   1 meggpavccq dpraelvery aaidvthlee adggpeptrn gvdppprara asvipgstsr  61 llparpslsa rklslqerpa gsyleaqagp yatgpashis prawrrptie shhvaisdae 121 dcvqlnqykl qseigkgayg vvrlaynese drhyamkvls kkkllkqygf prrppprgsq 181 aaqggpakql lplervyqei ailkkldhvn vvklievldd paednlylvf dllrkgpvme 241 vpcdkpfsee qarlylrdvi lgleylhcqk ivhrdikpsn lllgddghvk iadfgvsnqf 301 egndaqlsst agtpafmape aisdsgqsfs gkaldvwatg vtlycfvygk cpfiddfila 361 lhrkiknepv vfpeepeise elkdlilkml dknpetrigv pdiklhpwvt kngeeplpse 421 eehcsvvevt eeevknsvrl ipswttvilv ksmlrkrsfg npfepqarre ersmsapgnl 481 lvkegfgegg kspelpgvqe deaas

In some embodiments of the methods of the disclosure, the wild type human CAMKK1 gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_172206.1, transcript variant 2):

(SEQ ID NO: 55)    1 agcagaacag agtatgcaat ttgggaagct gtggtgtggc tgcagtggag agttcccaac   61 aaggctacgc agaagaaccc ccttgactga agcaatggag gggggtccag ctgtctgctg  121 ccaggatcct cgggcagagc tggtagaacg ggtggcagcc atcgatgtga ctcacttgga  181 ggaggcagat ggtggcccag agcctactag aaacggtgtg gaccccccac cacgggccag  241 agctgcctct gtgatccctg gcagtacttc aagactgctc ccagcccggc ctagcctctc  301 agccaggaag ctttccctac aggagcggcc agcaggaagc tatctggagg cgcaggctgg  361 gccttatgcc acggggcctg ccagccacat ctccccccgg gcctggcgga ggcccaccat  421 cgagtcccac cacgtggcca tctcagatgc agaggactgc gtgcagctga accagtacaa  481 gctgcagagt gagattggca agggtgccta cggtgtggtg aggctggcct acaacgaaag  541 tgaagacaga cactatgcaa tgaaagtcct ttccaaaaag aagttactga agcagtatgg  601 ctttccacgt cgccctcccc cgagagggtc ccaggctgcc cagggaggac cagccaagca  661 gctgctgccc ctggagcggg tgtaccagga gattgccatc ctgaagaagc tggaccacgt  721 gaatgtggtc aaactgatcg aggtcctgga tgacccagct gaggacaacc tctatttggt  781 gtttgacctc ctgagaaagg ggcccgtcat ggaagtgccc tgtgacaagc ccttctcgga  841 ggagcaagct cgcctctacc tgcgggacgt catcctgggc ctcgagtact tgcactgcca  901 gaagatcgtc cacagggaca tcaagccatc caacctgctc ctgggggatg atgggcacgt  961 gaagatcgcc gactttggcg tcagcaacca gtttgagggg aacgacgctc agctgtccag 1021 cacggcggga accccagcat tcatggcccc cgaggccatt tctgattccg gccagagctt 1081 cagtgggaag gccttggatg tatgggccac tggcgtcacg ttgtactgct ttgtctatgg 1141 gaagtgccca ttcatcgacg atttcatcct ggccctccac aggaagatca agaatgagcc 1201 cgtggtgttt cctgaggagc cagaaatcag cgaggagctc aaggacctga tcctgaagat 1261 gttagacaag aatcccgaga cgagaattgg ggtgccagac atcaagttgc acccttgggt 1321 gaccaagaac ggggaggagc cccttccttc ggaggaggag cactgcagcg tggtggaggt 1381 gacagaggag gaggttaaga actcagtcag gctcatcccc agctggacca cggtgatcct 1441 ggtgaagtcc atgctgagga agcgttcctt tgggaacccg tttgagcccc aagcacggag 1501 ggaagagcga tccatgtctg ctccaggaaa cctactggtg aaagaagggt ttggtgaagg 1561 gggcaagagc ccagagctcc ccggcgtcca ggaagacgag gctgcatcct gagcccctgc 1621 atgcacccag ggccacccgg cagcacactc atcccgcgcc tccagaggcc cacccctcat 1681 gcaacagccg cccccgcagg cagggggctg gggactgcag ccccactccc gcccctcccc 1741 catcgtgctg catgacctcc acgcacgcac gtccagggac agactggaat gtatgtcatt 1801 tggggtcttg ggggcagggc tcccacgagg ccatcctcct cttcttggac ctccttggcc 1861 tgacccattc tgtggggaaa ccgggtgccc atggagcctc agaaatgcca cccggctggt 1921 tggcatggcc tggggcagga ggcagaggca ggagaccaag atggcaggtg gaggccaggc 1981 ttaccacaac ggaagagacc tcccgctggg gccgggcagg cctggctcag ctgccacagg 2041 catatggtgg agaggggggt accctgccca ccttggggtg gtggcaccag agctcttgtc 2101 tattcagacg ctggtatggg ggctcggacc cctcactggg gacagggcca gtgttggaga 2161 attctgattc cttttttgtt gtcttttact tttgttttta acctgggggt tcggggagag 2221 gccctgcttg ggaacatctc acgagctttc ctacatcttc cgtggttccc agcacagccc 2281 aagattattt ggcagccaag tggatggaac taactttcct ggactgtgtt tcgcattcgg 2341 cgttatctgg aaagtggact gaacggaatc aagctctgag cagaggcctg aagcggaagc 2401 accacatcgt ccctgcccat ctcactctct cccttgatga tgcccctaga gctgaggctg 2461 gagaagacac cagggctgac tttgaccgag ggccatggac gcgacaggcc tgtggccctg 2521 cgcatgctga aataactgga acccagcctc tcctcctaca ccggcctacc catctgggcc 2581 caagagctgc actcacactc ctacaacgaa ggacaaactg tccaggtcgg agggatcacg 2641 agacacagaa cctggagggg tgtgcacgct ggcaggtggc ctctgcggca attgcctcac 2701 cctgaggaca tcagcagtca gcctgctcag agcgggggtg ctggagcgcg tgcagacaca 2761 gctcttccgg agcagccttc accttctctc tgggatcagt gtccggctgg ccgacgtggc 2821 atttgctgac cgaatgctca tagaggttga cccccacagg gtcacgcagg actcggacac 2881 tgccctggaa acatggatgg acaagggctt ttggccacag gtgtgggtgt cctgttggag 2941 gagggcttgt ttggagaagg gaggctggct gggggagaaa cccggatccc gctgcatctc 3001 cgcgcctgtg ggtgcatgtc gcgtgctcat ctgttgcaca cagctcactc gtatgtcctg 3061 cactggtaca tgcatctgta atacagtttc tacgtctatt taaggctagg agccgaatgt 3121 gccccattgt cagtgggtcc acgtttctcc ccggctcctc tgggctaagg cagtgtggcc 3181 cgaagcttaa aaagttactc ggtactgttt ttaagaacac ttttatagag ttagtggaag 3241 gcaagttaag agccaatcac tgatccccaa gtgtttcttg agcatctggt ctggggggac 3301 cactttgatc ggacccaccc ttggaaagct caggggtagg cccaggtggg atgctcaccc 3361 tgtcactgag ggttttggtt ggcatcgttg tttttgaatg tagcacaagc gatgagcaaa 3421 ctctataaga gtgttttaaa aattaacttc ccaggaagtg agttaaaaac aataaaagcc 3481 ctttcttgag ttaaaaagaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaa

In some embodiments of the methods of the disclosure, the wild type human CAMKK1 gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_757343.2, transcript variant 2):

(SEQ ID NO: 56)   1 mqfgklwcgc sgefptrlrr rtplteameg gpavccqdpr aelvervaai dvthleeadg  61 gpeptrngvd pppraraasv ipgstsrllp arpslsarkl slgerpagsy leaqagpyat 121 gpashispra wrrptieshh vaisdaedcv qlnqyklqse igkgaygvvr laynesedrh 181 yamkvlskkk llkqygfprr ppprgsqaaq ggpakqllpl ervyqeiail kkldhvnvvk 241 lievlddpae dnlylvfdll rkgpvmevpc dkpfseeqar lylrdvilgl eylhcqkivh 301 rdikpsnlll gddghvkiad fgvsnqfegn daqlsstagt pafmapeais dsgqsfsgka 361 ldvwatgvtl ycfvygkcpf iddfilalhr kiknepvvfp eepeiseelk dlilkmldkn 421 petrigvpdi klhpwvtkng eeplpseeeh csvvevteee vknsvrlips wttvilvksm 481 lrkrsfgnpf epqarreers msapgnllvk egfgeggksp elpgvqedea as

In some embodiments of the methods of the disclosure, the wild type human CAMKK1 gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_172207.2, transcript variant 3):

(SEQ ID NO: 57)    1 ctgggcccca gcgaggcggt ggggcggggc ggggcggggc ggggcgcgca gcaggagcga   61 gtggggccgc ccgccgggcc gcggacactg tcgcccggcg cccaggttcc caacaaggct  121 acgcagaaga acccccttga ctgaagcaat ggaggggggt ccagctgtct gctgccagga  181 tcctcgggca gagctggtag aacgggtggc agccatcgat gtgactcact tggaggaggc  241 agatggtggc ccagagccta ctagaaacgg tgtggacccc ccaccacggg ccagagctgc  301 ctctgtgatc cctggcagta cttcaagact gctcccagcc cggcctagcc tctcagccag  361 gaagctttcc ctacaggagc ggccagcagg aagctatctg gaggcgcagg ctgggcctta  421 tgccacgggg cctgccagcc acatctcccc ccgggcctgg cggaggccca ccatcgagtc  481 ccaccacgtg gccatctcag atgcagagga ctgcgtgcag ctgaaccagt acaagctgca  541 gagtgagatt ggcaagggtg cctacggtgt ggtgaggctg gcctacaacg aaagtgaaga  601 cagacactat gcaatgaaag tcctttccaa aaagaagtta ctgaagcagt atggctttcc  661 acgtcgccct cccccgagag ggtcccaggc tgcccaggga ggaccagcca agcagctgct  721 gcccctggag cgggtgtacc aggagattgc catcctgaag aagctggacc acgtgaatgt  781 ggtcaaactg atcgaggtcc tggatgaccc agctgaggac aacctctatt tggccctgca  841 gaaccaggcc cagaatatcc agttagattc aacaaatatc gccaagcccc actccctgct  901 tccctctgag cagcaagaca gtggatccac gtgggctgcg cgctcagtgt ttgacctcct  961 gagaaagggg cccgtcatgg aagtgccctg tgacaagccc ttctcggagg agcaagctcg 1021 cctctacctg cgggacgtca tcctgggcct cgagtacttg cactgccaga agatcgtcca 1081 cagggacatc aagccatcca acctgctcct gggggatgat gggcacgtga agatcgccga 1141 ctttggcgtc agcaaccagt ttgaggggaa cgacgctcag ctgtccagca cggcgggaac 1201 cccagcattc atggcccccg aggccatttc tgattccggc cagagcttca gtgggaaggc 1261 cttggatgta tgggccactg gcgtcacgtt gtactgcttt gtctatggga agtgcccatt 1321 catcgacgat ttcatcctgg ccctccacag gaagatcaag aatgagcccg tggtgtttcc 1381 tgaggagcca gaaatcagcg aggagctcaa ggacctgatc ctgaagatgt tagacaagaa 1441 tcccgagacg agaattgggg tgccagacat caagttgcac ccttgggtga ccaagaacgg 1501 ggaggagccc cttccttcgg aggaggagca ctgcagcgtg gtggaggtga cagaggagga 1561 ggttaagaac tcagtcaggc tcatccccag ctggaccacg gtgatcctgg tgaagtccat 1621 gctgaggaag cgttcctttg ggaacccgtt tgagccccaa gcacggaggg aagagcgatc 1681 catgtctgct ccaggaaacc tactggtgta agtactggtg ggccagggac tgccgggcac 1741 tccctggagt tgggtgggga ggtctgaggc ccatcctccc actctcactg tcgttgggcc 1801 aaggccagag cctggggact tggccaggtc tcggtgttgg ccccatttgc atctctgtcc 1861 ccaaggttag tcggggctag aagggacctt ttgggcccag ctcttgcttc attcctgggg 1921 ccagcatccc tcacacacac acttccaggg atgaggagct cacgcagccc ctccatggga 1981 caggaagacc cttcttccat gcagcttgat gtcactctct cactgggtcc agcccctctg 2041 gggcttcaaa tctgtggccc cctcagccct tggcagcctg gcagaggttt gcagacaggc 2101 tgatgttggc ttcctgtagg aggctggcgg gctgtagagg aggggtgctg gcccctctgc 2161 ctggccctgg ggactgttgg ctgctctccc aagtggccca ggctgcctgc agccattgct 2221 ggggctctgt gcccagtcag cactttgtga gtgcttgttc agtgagtaag cagggacagg 2281 ctggccggtg gaccacggga gaggaacccg cattggccga gggctcccta tggtgagcca 2341 cgcctgtggg ttcaccacct cctaggaggg tccagaaaag cagctcccca agcctgtgcg 2401 cctcgtcctc agcagatcca ccttcttcac tataataaaa gccagtctgg gatgctaaaa 2461 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 2521 aaaaaaaaaa aaaaa

In some embodiments of the methods of the disclosure, the wild type human CAMKK1 gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_757344.2, transcript variant 3):

(SEQ ID NO: 58)   1 meggpavccq dpraelvery aaidvthlee adggpeptrn gvdppprara asvipgstsr  61 llparpslsa rklslqerpa gsyleaqagp yatgpashis prawrrptie shhvaisdae 121 dcvqlnqykl qseigkgayg vvrlaynese drhyamkvls kkkllkqygf prrppprgsq 181 aaqggpakql lplervyqei ailkkldhvn vvklievldd paednlylal qnqaqniqld 241 stniakphsl lpseqqdsgs twaarsvfdl lrkgpvmevp cdkpfseeqa rlylrdvilg 301 leylhcqkiv hrdikpsnll lgddghvkia dfgvsnqfeg ndaqlsstag tpafmapeai 361 sdsgqsfsgk aldvwatgvt lycfvygkcp fiddfilalh rkiknepvvf peepeiseel 421 kdlilkmldk npetrigvpd iklhpwvtkn geeplpseee hcsvvevtee evknsvrlip 481 swttvilvks mlrkrsfgnp fepqarreer smsapgnllv

In some embodiments of the methods of the disclosure, the wild type human MMPI gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NM_002423.4):

(SEQ ID NO: 59)    1 gaaaacacca aatcaaccat aggtccaaga acaattgtct ctggacggca gctatgcgac   61 tcaccgtgct gtgtgctgtg tgcctgctgc ctggcagcct ggccctgccg ctgcctcagg  121 aggcgggagg catgagtgag ctacagtggg aacaggctca ggactatctc aagagatttt  181 atctctatga ctcagaaaca aaaaatgcca acagtttaga agccaaactc aaggagatgc  241 aaaaattctt tggcctacct ataactggaa tgttaaactc ccgcgtcata gaaataatgc  301 agaagcccag atgtggagtg ccagatgttg cagaatactc actatttcca aatagcccaa  361 aatggacttc caaagtggtc acctacagga tcgtatcata tactcgagac ttaccgcata  421 ttacagtgga tcgattagtg tcaaaggctt taaacatgtg gggcaaagag atccccctgc  481 atttcaggaa agttgtatgg ggaactgctg acatcatgat tggctttgcg cgaggagctc  541 atggggactc ctacccattt gatgggccag gaaacacgct ggctcatgcc tttgcgcctg  601 ggacaggtct cggaggagat gctcacttcg atgaggatga acgctggacg gatggtagca  661 gtctagggat taacttcctg tatgctgcaa ctcatgaact tggccattct ttgggtatgg  721 gacattcctc tgatcctaat gcagtgatgt atccaaccta tggaaatgga gatccccaaa  781 attttaaact ttcccaggat gatattaaag gcattcagaa actatatgga aagagaagta  841 attcaagaaa gaaatagaaa cttcaggcag aacatccatt cattcattca ttggattgta  901 tatcattgtt gcacaatcag aattgataag cactgttcct ccactccatt tagcaattat  961 gtcacccttt tttattgcag ttggtttttg aatgtctttc actcctttta aggataaact 1021 cctttatggt gtgactgtgt cttattcatc tatacttgca gtgggtagat gtcaataaat 1081 gttacataca caaataaata aaatgtttat tccatggtaa atttaaaaaa aaaaaaaaaa 1141 aaaaaaaaaa aaa

In some embodiments of the methods of the disclosure, the wild type human MMPI gene of the disclosure consists of or comprises the amino acid sequence (Genbank Accession number: NP_002414.1):

(SEQ ID NO: 60)   1 mrltvlcavc llpgslalpl pqeaggmsel qweqaqdylk rfylydsetk nansleaklk  61 emqkffglpi tgmlnsrvie imqkprcgvp dvaeyslfpn spkwtskvvt yrivsytrdl 121 phitvdrlvs kalnmwgkei plhfrkvvwg tadimigfar gahgdsypfd gpgntlahaf 181 apgtglggda hfdederwtd gsslginfly aathelghsl gmghssdpna vmyptygngd 241 pqnfklsqdd ikgiqklygk rsnsrkk

In some embodiments of the methods of the disclosure, the wild type human TERC gene of the disclosure consists of or comprises the nucleic acid sequence (Genbank Accession number: NR_001566.1):

(SEQ ID NO: 61)   1  gggttgcgga gggtgggcct gggaggggtg gtggccattt tttgtctaac cctaactgag  61 aagggcgtag gcgccgtgct tttgctcccc gcgcgctgtt tttctcgctg actttcagcg 121 ggcggaaaag cctcggcctg ccgccttcca ccgttcattc tagagcaaac aaaaaatgtc 181 agctgctggc ccgttcgccc ctcccgggga cctgcggcgg gtcgcctgcc cagcccccga 241 accccgcctg gaggccgcgg tcggcccggg gcttctccgg aggcacccac tgccaccgcg 301 aagagttggg ctctgtcagc cgcgggtctc tcgggggcga gggcgaggtt caggcctttc 361 aggccgcagg aagaggaacg gagcgagtcc ccgcgcgcgg cgcgattccc tgagctgtgg 421 gacgtgcacc caggactcgg ctcacacatg c

Definitions

The following definitions are included for the purpose of understanding the present subject matter and for constructing the appended patent claims. Abbreviations used herein have their conventional meaning within the chemical and biological arts.

Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art. Any methods, devices and materials similar or equivalent to those described herein can be used in the practice of this disclosure. The following definitions are provided to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present disclosure.

As used herein, the term “FILD” refers to fibrotic interstitial lung disease.

As used herein, the term “FIP” refers to Familial Interstitial Pneumonia.

As used herein, the term “HRCT” refers to high-resolution CT (HRCT).

As used herein, the term “ILA” refers to asymptomatic interstitial lung abnormalities.

As used herein, the term “IPF” refers to idiopathic pulmonary fibrosis.

As used herein, the term “PBMC” refers to peripheral blood mononuclear cell.

As used herein, the term “alleviate” is meant to describe a process by which the severity of a sign or symptom of a disorder is decreased. Importantly, a sign or symptom can be alleviated without being eliminated. In a preferred embodiment, the administration of pharmaceutical compositions disclosed herein leads to the elimination of a sign or symptom, however, elimination is not required. Effective dosages are expected to decrease the severity of a sign or symptom. A sign is an objective indication of a medical condition that is observable or detectable by a medical professional or lay person (e.g. family member) (for example, with respect to fibrotic pulmonary disease, signs include, but are not limited to, changes in body weight, changes in body temperature and the presence of a fibrotic lesion in one or both lungs detectable by radiography).

A symptom is an indication of disease that may be a sign but may also be exclusively observable or subjectively experienced by the subject (for example, with respect to fibrotic pulmonary disease, symptoms may include but are not limited to, a dry or hacking cough, a sore throat, a tight chest, shortness of breath, and a feeling of exhaustion or malaise).

In one aspect, the terms “co-administered” and “co-administration” as relating to a subject refer to administering to the subject a compound of the invention or salt thereof along with a compound that may also treat the disorders or diseases contemplated within the invention. In one embodiment, the co-administered compounds are administered separately, or in any kind of combination as part of a single therapeutic approach. The co-administered compound may be formulated in any kind of combinations as mixtures of solids and liquids under a variety of solid, gel, and liquid formulations, and as a solution.

As used herein, the term “composition” or “pharmaceutical composition” refers to a mixture of at least one compound useful within the disclosure with a pharmaceutically acceptable carrier. The pharmaceutical composition facilitates administration of the compound to a patient or subject. Multiple techniques of administering a compound exist in the art including, but not limited to, intravenous, oral, aerosol, parenteral, ophthalmic, nasal, pulmonary and topical administration.

A “disease” as used herein is a state of health of an animal or subject wherein the animal or subject cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal's or subject's health continues to deteriorate.

A “disorder” as used herein in an animal is a state of health in which the animal or subject is able to maintain homeostasis, but in which the animal's or subject's state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a further decrease in the animal's or subject's state of health.

As used herein, the terms “effective amount,” “pharmaceutically effective amount” and “therapeutically effective amount” refer to a nontoxic but sufficient amount of an agent to provide the desired biological result. That result may be reduction and/or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. An appropriate therapeutic amount in any individual case may be determined by one of ordinary skill in the art using routine experimentation.

As used herein, the term “fibrotic lung disease” or “fibroid lung disease” or “pulmonary fibrosis” or “scarring of the lung” refers to a group of diseases characterized by the formation or development of excess fibrous connective tissue (fibrosis) in the lungs. Symptoms of pulmonary fibrosis are mainly: shortness of breath, particularly with exertion; chronic dry, hacking coughing; fatigue and weakness; chest discomfort; and loss of appetite and rapid weight loss. Pulmonary fibrosis may be a secondary effect of other diseases, most of them being classified as interstitial lung diseases, such as autoimmune disorders, viral infections or other microscopic injuries to the lung. Pulmonary fibrosis can also appear without any known cause (“idiopathic”). Idiopathic pulmonary fibrosis is a diagnosis of exclusion of a characteristic set of histologic/pathologic features known as usual interstitial pneumonia (UIP).

Diseases and conditions that may cause pulmonary fibrosis as a secondary effect include: inhalation of environmental and occupational pollutants (asbestosis, silicosis and gas exposure); hypersensitivity pneumonitis, most often resulting from inhaling dust contaminated with bacterial, fungal, or animal products; cigarette smoking; connective tissue diseases such as rheumatoid arthritis, SLE; scleroderma, sarcoidosis and Wegener's granulomatosis; infections; medications such as amiodarone, bleomycin (pingyangmycin), busulfan, methotrexate, apomorphine and nitrofurantoin; and radiation therapy to the chest.

As used herein, a “subject in need thereof” is a subject suffering from fibrotic lung disease relative to the population at large. For example, the subject is a patient who is or is about to be administered with comprising administering to the subject an effective amount of a therapeutic agent. For example, the subject is asymptomatic and is at risk of developing the fibrotic lung disease. A “subject” includes a mammal. The mammal can be e.g., any mammal, e.g., a human, primate, bird, mouse, rat, fowl, dog, cat, cow, horse, goat, camel, sheep or pig. Preferably, the mammal is a human.

As used herein, the term “pharmaceutically acceptable” refers to a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the compound, and is relatively non-toxic, i.e., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound useful within the invention within or to the patient such that it may perform its intended function. Typically, such constructs are carried or transported from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, including the compound useful within the invention, and not injurious to the patient. Some examples of materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations.

Pharmaceutically acceptable carriers of the disclosure include, but are not limited to, pharmaceutically acceptable materials, compositions or carriers, such as a liquid or solid fillers, stabilizers, dispersing agents, suspending agents, diluents, excipients, thickening agents, solvents or encapsulating materials, involved in carrying or transporting a compound useful within the invention within or to the patient such that it may perform its intended function. Typically, such constructs are carried or transported from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, including the compound useful within the invention, and not injurious to the patient. Some examples of materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations.

Suitable forms for administration include forms suitable for systemic administration, oral administration, for example by a capsule or tablet. Once formulated, the compositions of the disclosure can be administered directly to the subject.

The term “prevent,” “preventing” or “prevention,” as used herein, means avoiding or delaying the onset of symptoms associated with a disease or condition in a subject that has not developed such symptoms at the time the administering of an agent or compound commences.

Compounds and Compositions

In some embodiments, compounds known to be useful in treating pulmonary fibrosis are useful within the methods of the invention. Non-limiting examples of such compounds are pirfenidone (5-methyl-1-phenylpyridin-2-one, or a salt or solvate thereof) and nintedanib (methyl (3Z)-3-{[(4-{methyl[(4-methylpiperazin-1-yl)acetyl]amino}phenyl)amino](phenyl)methylidene}-2-oxo-2,3-dihydro-1 H-indole-6-carboxylate, or a salt or solvate thereof).

In some embodiments, the subject identified as having MUC5B promoter polymorphism rs35705950 is administered a compound contemplated within the disclosure. In some embodiments, the subject is a mammal. In other embodiments, the mammal is a human.

Administration/Dosage/Formulations

The regimen of administration may affect what constitutes an effective amount. The therapeutic formulations may be administered to the subject either prior to or after the onset of a disease or disorder contemplated in the invention. Further, several divided dosages, as well as staggered dosages may be administered daily or sequentially, or the dose may be continuously infused, or may be a bolus injection. Further, the dosages of the therapeutic formulations may be proportionally increased or decreased as indicated by the exigencies of the therapeutic or prophylactic situation.

Administration of the compositions of the present disclosure to a patient, preferably a mammal, more preferably a human, may be carried out using known procedures, at dosages and for periods of time effective to treat a disease or disorder contemplated in the invention. An effective amount of the therapeutic compound necessary to achieve a therapeutic effect may vary according to factors such as the state of the disease or disorder in the patient; the age, sex, and weight of the patient; and the ability of the therapeutic compound to treat a disease or disorder contemplated in the invention. Dosage regimens may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. A non-limiting example of an effective dose range for a therapeutic compound of the invention is from about 1 and 5,000 mg/kg of body weight/per day. One of ordinary skill in the art would be able to study the relevant factors and make the determination regarding the effective amount of the therapeutic compound without undue experimentation. Actual dosage levels of the active ingredients in the pharmaceutical compositions of this invention may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

The precise therapeutically effective amount for a human subject will depend upon the severity of the disease state, the general health of the subject, the age, weight and gender of the subject, diet, time and frequency of administration, drug combination(s), reaction sensitivities and tolerance/response to therapy. This amount can be determined by routine experimentation and is within the judgement of the clinician.

A medical doctor, e.g., physician or veterinarian, having ordinary skill in the art may readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the invention employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

A suitable dose of a compound of the disclosure may be in the range of from about 0.01 mg to about 5,000 mg per day, such as from about 0.1 mg to about 1,000 mg, for example, from about 1 mg to about 500 mg, such as about 5 mg to about 250 mg per day. The dose may be administered in a single dosage or in multiple dosages, for example from 1 to 4 or more times per day. When multiple dosages are used, the amount of each dosage may be the same or different. For example, a dose of 1 mg per day may be administered as two 0.5 mg doses, with about a 12-hour interval between doses.

In some embodiments of the methods of the disclosure, the therapeutic agent comprises pirfenidone. In some embodiments, the effective dosage is administered orally as a capsule or a tablet. In some embodiments, including those embodiments wherein the therapeutic agent comprises pirfenidone, the effective dosage is about 2400 mg/day. In some embodiments, the effective dosage is administered according to an escalating dosage regimen. In some embodiments, including those embodiments wherein the therapeutic agent comprises pirfenidone, the escalating dosage regimen comprises (a) administering to the subject about 800 mg of pirfenidone per day for a first week; (b) administering to the subject about 1600 mg of pirfenidone per day for a second week; and (c) administering to the subject about 2400 mg of pirfenidone per day for the remainder of the treatment. In some embodiments, including those embodiments wherein the therapeutic agent comprises pirfenidone, the escalating dosage regimen comprises (a) administering to the subject a capsule or tablet comprising about 250 mg of pirfenidone three times a day for a first week; (b) administering to the subject two capsules or tablets comprising about 250 mg of pirfenidone three times a day for a second week; and (c) administering to the subject three capsules or tablets comprising about 250 mg of pirfenidone three times a day for the remainder of the treatment. In some embodiments of the escalating dosage regimen, the capsule or tablet comprises 267 mg of pirfenidone.

In some embodiments of the methods of the disclosure, the therapeutic agent comprises nintedanib. In some embodiments, the effective dosage is administered orally as a capsule or a tablet. In some embodiments, including those embodiments wherein the therapeutic agent comprises nintedanib, the effective dosage is about 300 mg/day. In some embodiments, the effective dosage is about 150 mg administered twice per day, wherein the daily doses are administered about 12 hours apart from one another. In some embodiments, including those embodiments wherein the therapeutic agent comprises nintedanib, the effective dosage is about 200 mg/day. In some embodiments, the effective dosage is about 100 mg administered twice per day, wherein the daily doses are administered about 12 hours apart from one another. In some embodiments, including those embodiments wherein the therapeutic agent comprises nintedanib, the effective dosage is administered according to a modified or interrupted dosage regimen. In some embodiments, the modified or interrupted dosage regimen comprises (a) administering to the subject about 300 mg of nintedanib per day until the subject presents an elevated level of liver enzymes compared to a control level of liver enzymes; (b) administering to the subject about 200 mg of nintedanib per day until the subject presents the control level of liver enzymes; and (c) administering to the subject about 300 mg of nintedanib per day for the remainder of the treatment; wherein the control level of liver enzymes is a level detected in the subject prior to an initiation of the treatment. In some embodiments, including those embodiments wherein the therapeutic agent comprises nintedanib, the modified or interrupted regimen comprises (a) administering to the subject a capsule or tablet comprising about 150 mg of nintedanib twice per day until the subject presents an elevated level of liver enzymes compared to a control level of liver enzymes; (b) administering to the subject two capsules or tablets comprising about 100 mg twice per day until the subject presents an elevated level of liver enzymes compared to a control level of liver enzymes; and (c) administering to the subject a capsule or tablet comprising about 150 mg of nintedanib twice per day for the remainder of the treatment; wherein the control level of liver enzymes is a level detected in the subject prior to an initiation of the treatment.

In some embodiments, the compositions of the invention are formulated using one or more pharmaceutically acceptable excipients or carriers. In one embodiment, the pharmaceutical compositions of the invention comprise a therapeutically effective amount of a compound of the invention and a pharmaceutically acceptable carrier.

The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity may be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms may be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal and the like. In many cases, it is preferable to include isotonic agents, for example, sugars, sodium chloride, or polyalcohols such as mannitol and sorbitol, in the composition.

It is also noted that the term “comprising” is intended to be open and permits but does not require the inclusion of additional elements or steps. When the term “comprising” is used herein, the terms “consisting essentially of” and “consisting of” are thus also encompassed and disclosed. Throughout the description, where compositions or combinations are described as having, including, or comprising specific components or steps, it is contemplated that compositions or combinations also consist essentially of, or consist of, the recited components. Similarly, where methods or processes are described as having, including, or comprising specific process steps, the processes also consist essentially of, or consist of, the recited processing steps.

All publications and patent documents cited herein are incorporated herein by reference as if each such publication or document was specifically and individually indicated to be incorporated herein by reference.

EXAMPLES

In order that the invention disclosed herein may be more efficiently understood, examples are provided below. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting the invention in any manner.

Example 1: Genetic Background of Asymptomatic Siblings of FIP Subjects

Asymptomatic siblings (>50 years old) of patients with established FIP underwent HRCT scan of the chest. HRCT scans were assessed for FILD by blinded thoracic radiologists; when possible, specific radiographic patterns were identified. PBMCs RNA and DNA were isolated. Genotyping for rs35705950 and microarray analysis were performed (SurePrint G3 Human Gene Expression Microarray). Data were analyzed using Partek Genomics Suite and RStudio. Four-hundred eighty-eight FIP siblings from 271 families were evaluated, 25 HRCT scans were excluded due to technically inadequacy, leaving 463 to be interpreted. Of these, 19% (n=88) met criteria for FILD. A subset of the positive FILD scans (n=58) were evaluated for specific interstitial patterns: the predominant radiographic finding was Usual Interstitial Pneumonia (UIP), documented as possible (n=37), probable (n=6), or definite (n=5) in 82.8% of these cases. DNA was available for 443 subjects (358 without and 85 with FILD). The minor allele (T) frequency (MAF) of rs35705950 was higher among those with evidence of FILD (MAF=0.29) than among those with normal appearing HRCT scans (MAF=0.21, p=0.005). The rs35705950 variant was associated with the presence of FILD (OR=1.90, 95% CI 1.10-3.30, p=0.02), and FILD was associated with age (OR=1.09, 95% CI 1.06-1.12, p=7.24×10-9), male sex (OR=1.81, 95% CI 1.04-3.16, p=0.04), and history of smoking (OR=1.94, 95% CI 1.11-3.40, p=0.02). Microarray analysis on PBMC RNA from 40 subjects with FILD and 105 unaffected siblings revealed 1,272 differentially expressed genes (FDR<0.05, fold-change>2); hierarchical clustering performed on the top 194 differentially expressed probes illustrates segregation of FILD subjects from unaffected siblings (FIG. 1).

Example 2: Role of MUC5B in Pathogenesis of IPF

Common genetic variants play major and similar roles in the development of both familial and sporadic IPF (Table 3), indicating a similar etiology for familial and sporadic IPF. A common gain-of-function MUC5B promoter variant rs35705950 is a strong risk factor (genetic and otherwise), accounting for at least 30% of the total risk of developing IPF (10) confirmed in 10 independent studies, including a GWAS (OR for T (minor) allele=4.51; 95% CI=3.91-5.21; P=7.21×10-95); 3) rs35705950 may be used to identify individuals with PrePF and is predictive of radiographic progression of PrePF. MUC5B promoter variant rs35705950 is present in over 50% of non-Hispanic white (NHW) patients with IPF and is associated with unique biological and clinical IPF phenotypes. PrePF can be predicted using a combination of clinical risk factors, the MUC5B promoter variant rs35705950, and a panel of biomarkers.

TABLE 1 Common IPF risk variants identified by targeted sequencing of risk loci in 3,642 IPF cases and 4,442 unaffected controls Common Nearest Minor MAF in OR Aa vs AA OR aa vs AA Chrm Variant Gene Annotation^(a) Allele cases (95% CI) (95% CI) P^(b) 3q26 rs2293607 TERC 3′ UTR C 0.2999 1.30 (1.18-1.43) 1.79 (1.49-2.15) 9.11 × 10⁻¹³ 4q22 rs2609260 FAM13A Intronic C 0.2289 1.35 (1.22-1.50) 1.96 (1.56-2.47) 1.03 × 10⁻¹³ 5p15 rs4449583 TERT Intronic T 0.2641 0.68 (0.62-0.75) 0.46 (0.39-0.55) 2.67 × 10⁻²⁵ 6p24 rs2076295 DSP Intronic G 0.5428 1.27 (1.14-1.42) 2.08 (1.83-2.37) 1.11 × 10⁻²⁹ 7q22 rs6963345 ZKSCAN1 Intronic A 0.4444 1.35 (1.22-1.50) 1.73 (1.51-1.99) 1.89 × 10⁻¹⁵ 10q24 rs2488000 OBFC1 Intronic T 0.08 0.70 (0.62-0.79)^(c) 7.13 × 10⁻⁹  11p15 rs35705950 MUC5B Promoter T 0.3533 5.45 (4.91-6.06) 18.68 (13.34-6.17)  9.60 × 10⁻²⁹⁵ 13q34 rs1278769 AK025511 3′ UTR A 0.1996 0.77 (0.70-0.85) 0.69 (0.56-0.86) 7.48 × 10⁻⁸  15q15 rs35700143 IVD — C 0.4118 0.76 (0.68-0.84) 0.63 (0.55-0.71) 3.44 × 10⁻¹² 19p13 rs12610495 DPP9 Intronic G 0.3398 1.22 (1.11-1.35) 1.59 (1.36-1.87) 3.11 × 10⁻⁹  OR, odds ratio. The minor allele is defined as the minor allele in the combined case and control group. ^(a)Based on SNPDOC; ^(b)P value adjusted for sex; ^(c)OR resulting from dominant test.

MUC5B is predicted is involved in the pathogenesis of IPF. FIG. 5 shows that MUC5B promoter variant is associated with enhanced MUC5B expression in both unaffected subjects and in patients with IPF and in IPF, MUC5B message and protein are expressed in bronchoalveolar epithelia (FIG. 6) and honeycomb cysts. In mice, the concentration of Muc5b is directly related to the fibroproliferative response to bleomycin (FIG. 7), Muc5b protein is expressed in the injured lung following bleomycin challenge, and enhanced production of Muc5b in mice appears to initiate endoplasmic reticulum (ER) stress in peripheral airways (FIGS. 8 and 9). Preliminary studies, also show that mucociliary clearance is decreased in mice that over-express Muc5b (SFTPC-Muc5b^(Tg)) and in humans with IPF (FIG. 10).

Interstitial lung abnormalities on HRCT scans show asymptomatic relatives of patients with familial IPF and in the elderly. Similar to patients with IPF, interstitial lung abnormalities in asymptomatic subjects are associated with advanced age, cigarette smoking, reduced lung volume and decreased exercise tolerance. Moreover, the MUC5B promoter variant rs35705950 is associated with a higher prevalence of interstitial lung abnormalities on HRCT scan and is predictive of radiographic progression. Suggesting that interstitial lung abnormalities on HRCT scan are a precursor of IPF. However, interstitial lung abnormalities are not specific and include non-fibrotic and fibrotic HRCT defects, and consequently, the prevalence of interstitial lung abnormalities (>5% in the general population ≥50 years of age is orders of magnitude higher than IPF.

To address the non-specificity of interstitial lung abnormalities, a novel entity—Preclinical Pulmonary Fibrosis (PrePF) was used. PrePF is reported more frequently among smokers and in families with two or more cases of pulmonary fibrosis. In the Framingham population, data shows that PrePF is present in 1.8% of the general population ≥50 years of age (in contrast, interstitial lung abnormalities were seen in 6.7%) and that the MUC5B promoter variant rs35705950 is predictive of those with PrePF (OR=6.3 per allele [95% CI 3.1-12.7). As shown herein, among asymptomatic first-degree family members of familial interstitial pneumonia (FIP) 14% have fibrotic interstitial changes on CT scan and 35% have interstitial abnormalities on transbronchial biopsy. Moreover, in the Framingham population, it is shown that rs35705950 is predictive of radiographic progression of PrePF (OR=2.8 per allele [95% CI 1.8-4.4]) which is associated with a greater FVC decline (P=0.0001) and an increased risk of death (HR=3.7 [95% CI 1.3, 10.7]; P=0.02), indicating that in addition to having radiographic features of IPF, PrePF has similar risk factors (age, gender, smoking, and MUC5B variant) and a progressive clinical course. While the MUC5B promoter variant is predictive of PrePF, rs35705950 is present in ≈19% (minor allele frequency (MAF)=0.09) of the NHW population, however IPF occurs infrequently (<0.1%). Thus, additional biomarkers may be used in combination with rs35705950 identify PrePF within at-risk populations.

The data provided herein suggest that 1) IPF is under-diagnosed; 2) PrePF is prevalent in at-risk populations; 3) approximately 75% of the cases of PrePF are progressive; 4) radiographic progression of PrePF is associated with increased morbidity and mortality; and 5) MUC5B variant rs35705950, peripheral blood biomarkers, clinical/biological, and radiographic screening should be useful in identifying those with PrePF (FIG. 11). While IPF takes years to develop, most patients with IPF are diagnosed in the advanced stage when little can be done to influence survival. Once the lung has undergone remodeling, the non-compliant, stiff lung matrix causes additional remodeling through activation of myofibroblasts, resulting in a feed-forward loop of lung remodeling. Earlier diagnosis of IPF detects subjects with a lower burden of fibrotic lung disease.

This disclosure provides a strategic approach to screening for early forms of IPF needs to be established (FIG. 11). While the MUC5B promoter variant is predictive of PrePF (defined as chest HRCT consistent with probable or definite fibrosis (e.g., bilateral subpleural reticular changes, honeycombing, or traction bronchiectasis) occurring in asymptomatic subjects ≥40 years of age that emerge from at-risk populations), the MUC5B promoter variant is present in ≈19% of the NHW population and IPF occurs infrequently (<0.1%). To study at-risk populations (asymptomatic siblings ≥40 years of age of patients with family or sporadic IPF), identification of genetic variants and biomarkers that increase the yield of patients with PrePF are used to establish screening tools and approaches that identify early stages of IPF. This approach changes the way IPF is diagnosed and treated, and is critical to developing interventions to prevent PrePF progression to established IPF. The methods provided in this disclosure fundmentally alter the clinical approach to patients with IPF from palliative to preventive (FIG. 10).

Example 3: Predictive Biomarker Profile for Established IPF

To address the development of a peripheral blood biomarker profile for IPF, an assay of the expression levels of >3700 plasma proteins was performed on plasma from 70 patients with established IPF and 70 controls. After controlling for multiple comparisons and appropriate co-variables, 57 proteins were up-regulated >1.5-fold (including surfactant proteins, MMPI, and C3) in the plasma of patients with IPF and 12 were significantly down-regulated (FIG. 2).

Example 4: Predictive Biomarker Profile for Early IPF

To evaluate a predictive biomarker profile in cases of preclinical pulmonary fibrosis (PrePF) derived from families with familial IPF (≥2 cases of IPF in a family), HRCT scans were performed on 496 asymptomatic family members ≥40 years of age previously phenotyped as unaffected from 263 families with familial IPF. PrePF, consistent with the operational definition (defined as abnormalities on chest HRCT consistent with probable or definite fibrosis (e.g., bilateral subpleural reticular changes, honeycombing, or traction bronchiectasis) occurring in asymptomatic subjects ≥40 years that emerge from at-risk populations), was present in 77 (15.5%) of 496 asymptomatic individuals from families with familial IPF. The minor allele frequency (MAF) of the MUC5B promoter variant was 0.29 in those with PrePF versus 0.21 in those without fibrosis (P=0.025). Preliminary analysis of PBMC gene expression profiles evaluated by microarrays from 38 cases of PrePF and 187 subjects without fibrosis identified 16 genes significantly differentially expressed between the two groups (p-value <0.05 and >1.5 fold change). Among genes differentially expressed in PrePF are those involved in innate immunity and inflammatory responses (SIGLEC14), antibacterial effects (ADM2), growth and motility (TSPAN5), and protein phosyphorylation (CAMKK1). Moreover, PBMC gene expression appears to contribute to the ability to predict PrePF in an at-risk population (FIG. 3).

Additionally, RNA-sequencing analysis was performed on 40 PrePF subjects and 80 subjects with a normal HRCT scan. Sequencing of the polyA-enriched libraries was prepared using Illumina TrueSEQ reagents and multiplexing 10 samples on each lane of HiSEQ4000 to obtain on average 35-40 million reads per sample. This high coverage allows for the consideration of a broad dynamic range of mRNA transcripts for biomarker selection. Platform selection of serum and plasma samples from the same subjects are used for proteomic analysis.

Example 5: Biomarker Identification

To examine for association between each biomarkers and PrePF, a multivariable logistic regression model for PrePF with biomarkers and covariates is used for inclusion and a step-wise forward selection procedure is constructed. Variables stay in the model if associated at P≤0.01 after adjustment for the variables already in the model. Protein biomarkers that are significantly associated with established IPF and the top 20 differentially expressed genes in PrePF are considered for inclusion in a multivariable model. The number of potential biomarkers allowed in the joint model is restricted to approximately 20 given the number cases of PrePF expected. Secondarily, interactions between MUC5B genotype and the other biomarkers are tested for, which allow for the possibility that different biomarker profiles are diagnostic in IPF patients with/without the MUC5B risk allele.

Example 6: Predictive Ability of Biomarkers

To test the predictive value of the combination of biomarkers associated with PrePF, the observed expression and other biomarker values from those associated with PrePF in the siblings of FIP patients is used to obtain the probability, for each sibling, having PrePF.

Following, a construct receiver operating characteristic (ROC) curves (see M. S. Pepe et al., Phases of biomarker development for early detection of cancer. Journal of the National Cancer Institute 93, 1054-1061 (2001)), is used to choose the probability threshold that maximizes the area under the ROC curve. This probability threshold is used to classify each individual as predicted to have PrePF or not, allowing calculation of the sensitivity, specificity, positive predictive value, and negative predictive value of the predictive model. The properties of the predictive model(s) in the independent set of siblings of patients with IPF are evaluated. Different aliquots are run for 10 samples for each assay at each time the assays is run in order to use those 10 samples to evaluate the need for standardization of the absolute values for each assay over time. Either the raw or standardized values, for a given model, is used to observe biomarker values in the PrePF siblings and non-PrePF siblings to obtain the probability of being in the disease group based on the model parameters developed using the FIP siblings. The thresholds identified among the FIP siblings are used to classify each individual as predicted to have PrePF or not. This categorization allows for the calculation of the sensitivity, specificity, positive predictive value, and negative predictive value of the predictive model among the siblings of independent cases of IPF to that observed in the siblings of FIP cases.

Power is calculated to detect differences between those with and without PrePF assuming 500 siblings and 10% (N=50) with PrePF. Assuming α=0.00005 (conservatively correcting for up to 1000 independent tests), we have 80% (90%) power to detect differences in protein or expression level of 0.74 (0.80) standard deviation between PrePF and unaffected siblings. These differences are larger than previously-observed protein and gene-expression levels in IPF patients and controls (see I. V. Yang et al., The peripheral blood transcriptome identifies the presence and extent of disease in idiopathic pulmonary fibrosis. PLoS One 7, e37708 (2012). With 50 PrePF and 450 unaffected, there is 90% power to bound the sensitivity of the biomarker-based classification of PrePF with a margin of error of 11% if the sensitivity is 65%, and 6.5% if the sensitivity is 95%; the margins of error for 65% and 95% sensitivity are 4.5% and 2.5%, respectively.

Example 7: MUC5B Promoter Variant r35705950is a Risk Factor for Rheumatoid Arthritis—Interstitial Lung Disease Methods Study Cohorts

This study included a discovery cohort and multi-ethnic replication cohorts. The discovery cohort included patients with RA, with and without ILD (RA-noILD) as assessed by chest HRCT, and controls, from the French RA-ILD network. The multi-ethnic replication cohorts were obtained from six countries (China, Greece, Japan, Mexico, the Netherlands and United States). This included patients with RA-ILD and RA-noILD patients, and controls. All cases fulfilled the 2010 European League Against Rheumatism-American College of Rheumatology (EULAR-ACR) and/or 1987 ACR revised criteria for RA. The ILD status of patients with RA was established by chest HRCT images that were centrally reviewed by experienced readers for each participating cohort. There was one cohort, the RA-noILD cases from the USA1 cohort, which was determined by self-report. The chest HRCT ILD pattern was classified as UIP, possible UIP or inconsistent with UIP according to international criteria and all readers were blinded to the clinical and genetic data. The institutional review boards at each institution approved all protocols, and all patients provided written informed consent.

Genotyping

Genotyping of the MUC5B rs35705950 single nucleotide polymorphism (SNP) involved use of Taqman Genotyping Assays (Applied Biosystems, Foster City, Calif., USA) as previously reported, by direct Sanger Sequencing or imputation from genome-wide association study data.

The additional common IPF risk variants on 3q26, 4q22, 5p15, 6p21.3, 6p24, 7q22, 10q24, 11p15.5, 13q34, 15q14-15, and 19p13 were genotyped by Taqman qPCR (Thermo Fisher Scientific, California) per the manufacturer's instructions.

Lung Tissue Analysis

In order to determine if MUC5B was expressed in RA-ILD ling tissue, we analyzed lung tissue was analyzed from nine patients with RA-ILD undergoing lung transplantation (University of California, San Francisco) compared to six unaffected controls with ILD (NHLBI Lung Tissue Research Consortium; https://ltrcpublic.com) or concordant expression of other relevant markers of pulmonary fibrosis. The tissue was formalin fixed, paraffin embedded and cut in 4 um sections. Tissue sections were deparaffinized in xylene, followed by dehydration in series of ethanol. Following citrate buffer antigen retrieval, slides were incubated overnight with primary antibodies against MUC5B (1:4000, Santa Cruz, Dallas, Tex.). Secondary antibody diluted 1:1000 tagged with HRP (Life Technologies) was visualized using an Aperio CS2 slide scanner (Leica, Buffalo Grove, Ill.).

Results Study Cohorts

This case-control genetic study included 620 RA-ILD cases, 614 RA-noILD cases and 5448 unaffected controls. The discovery cohort included 118 RA-ILD cases, 105 RAnoILD cases and 1229 unaffected controls. The multi-ethnic replication sample included 502 RA-ILD, 509 RA-noILD cases and 4219 unaffected controls.

Characteristics of the Discovery Cohort

As compared with RA-noILD, patients with RA-ILD were more frequently male, older and more frequently smoked cigarettes (54.7% versus 36.1%) (FIG. 13). However, after adjusting for sex, the relationship between RA-ILD and cigarette smoking was no longer statistically significant (FIG. 13). After adjustment, RA-ILD and RA-noILD patients did not differ in rheumatoid factor (RF) and/or anti-citrullinated protein antibody (ACPA) positivity, erosive status of RA, exposure to methotrexate or the mean RA duration from diagnosis at inclusion in the cohort. Overall, 41% of patients with RA-ILD had a UIP or possible UIP HRCT pattern.

MUC5B Promoter Variant and Risk of Rheumatoid Arthritis-Associated Interstitial Lung Disease

Comparison of RA-noILD and controls revealed that none of the cohorts (discovery cohort and multi-ethnic cohorts) demonstrated a significant difference in the frequency of the MUC5B promoter variant (FIG. 14; FIG. 16A), suggesting a lack of association between the MUC5B promoter variant and RA. In the discovery cohort, the minor allele frequency (MAF) of the MUC5B promoter variant was 10.9% in unaffected controls and 32.6% in cases of RAILD; this variant was in Hardy-Weinberg equilibrium (HWE) in both study groups. I In the discovery population, after controlling for sex we detected a significant association between the MUC5B promoter variant and RA-ILD when compared to non-RA controls (ORadj=3.8; 95% CI, 2.8 to 5.2; P=9.7×10-17) (FIG. 14). Similar to the discovery population, the MUC5B promoter variant was significantly over-represented among the

cases of RA-ILD compared to unaffected non-RA controls in all of the multi-ethnic study case series, except in the two Asian case series (FIG. 14). Given that the MUC5B promoter variant is under-represented in Asian populations compared to non-Hispanic whites (FIG. 14; www.ncbi.nlm.nih.gov/projects/SNP/snp_refcgi?rs=35705950), a likely explanation, especially given the consistent point estimates, for the absence of a significant relationship between the MUC5B promoter variant and RA-ILD is that the analysis of the two Asian case series is likely underpowered. The relationship between the MUC5B promoter variant and RA-ILD in combined multi-ethnic study case series (ORadj=4.7; 95% CI, 3.9 to 5.8; P=1.3×10-49) (FIG. 14) (FIG. 16B) validated the observed association between the MUC5B promoter variant and RA-ILD in the discovery study population.n addition, the cases of RA-ILD in the study populations from Greece and USA-1 were not in HWE, suggesting (as has been observed in cases of IPF 14), that the MUC5B promoter variant and/or common variants in high or complete linkage disequilibrium with the MUC5B promoter variant should be considered as causative in these cases of RA-ILD. For the comparison with non-RA controls, the best-fitting genetic model for the three study populations (discovery population, combined multi-ethnic case series, and combined analysis) for the association of the MUC5B MUC5B RS35705950 AND RISK OF ITNERSTITIAL LUNG DISEASE AMONG PATIENTS WITH RHEUMATOID ARTHRITIS

To further investigate whether the MUC5B promoter variant rs35705950 contributes to the risk of ILD among patients with RA, we compared RA-ILD and RA-noILD patients, adjusting for sex, age at inclusion and cigarette smoking. In the discovery cohort, the MUC5B variant was associated with RA-ILD (ORadj=3.1; 95% CI, 1.6 to 6.3; P=9.4×10⁻⁴), and this finding was replicated in the aggregate multi-ethnic cohort (ORadj=2.9; 95% CI, 1.1 to 8.4; P=0.04) and the combined analysis (ORadj, 3.1; 95% CI, 1.8 to 5.4; P=7.4×10⁻⁵) (FIG. 14; FIG. 16C). For the comparison of RA-ILD with RA-noILD, the best-fitting genetic model for the three study cohorts (discovery population, combined multi-ethnic case series, and combined analysis) was dominant. After adjusting for covariates, no association between tobacco smoking and the risk of ILD among patients with RA was found and no interaction of tobacco smoke exposure with the MUC5B promoter variant was observed (ORadj=0.7; 95% CI, 0.3 to 1.9; P=0.51).

MUC5B rs35705950 and UIP on HRCT Scan

Limiting the RA-ILD cases to those with radiographic evidence of definite or possible UIP on HRCT scan, the association observed in the discovery cohort (ORadj=5.0; 95% CI, 2.1 to 12.3; P=3.0×10⁻⁴), was replicated in the combined multi-ethnic cohort (ORadj=9.2; 95% CI, 2.3 to 38.7; P=1.8×10⁻³) (FIG. 16C), and was observed in the combined cohort analysis (ORadj=6.1; 95% CI, 2.9 to 13.1; P=2.5×10⁻⁶) (FIG. 16C). In the combined analysis, the comparison of odds ratios for UIP RA-ILD vs RA-noILD (ORadj=6.1; 95% CI, 2.9 to 13.1; P=2.5×10⁻⁶) to non-UIP RA-ILD vs RA-noILD (ORadj=1.3; 95% CI, 0.6 to 2.8; P=0.46) was statistically significant (P=0.02), suggesting that the effect of the MUC5B promoter variant was restricted to the UIP RA-ILD sub-phenotype (FIG. 16C). Finally, consistent with our previous findings, the MUC5B promoter variant was found to increase the risk of developing a UIP pattern among patients with RA-ILD through a dominant model in the discovery, replication and combined analysis; the odds of having a UIP and possible UIP pattern for patients with RA-ILD carrying at least one MUC5B rs35705950 T risk allele were 2.9 times greater than individuals having the GG genotype (ORadj=2.9; 95% CI, 1.7 to 4.8; P=5.1×10-5) (FIG. 15; FIG. 16C). After adjusting for covariates, tobacco smoking exposure did not contribute to a specific HRCT pattern for RA-ILD and no interaction with the MUC5B rs35705950 variant was detected.

Sites of MUC5B Expression in RA-ILD

We performed immunohistochemical staining for MUC5B in nine RA-ILD lung tissue explants (5 GG and 4 GT) and 6 unaffected controls (3 GG and 3 GT). Similar to what has been reported in IPF, RA-ILD lung tissue demonstrated MUC5B in the cytoplasm of the bronchioles and in areas of microscopic honeycombing, including staining of the metaplastic epithelia lining the honeycomb cysts and the mucous within the cyst (FIG. 17). The controls demonstrated MUC5B expression in the bronchioles only. There were no obvious differences in MUC5B expression by genotype.

Exploratory Genetic Association Study of 12 Common IPF Risk Variants in RA-ILD

Having provided evidence for the contribution of the dominant IPF genetic risk variant, i.e. the MUC5B promoter variant, to RA-ILD, we decided to test the association of 12 additional common IPF risk variants with RA-ILD (FIG. 29). This exploratory study included 272 RA-ILD and 242 RA-noILD patients from the France, USA-1 and Mexico case series. Taking into account the relatively small sample size and related low power of detection corresponding P-values, Odds Ratio and 95% CI for the 12 candidate variants were considered as descriptive and Bonferoni correction was therefore not applied (Table 4). Comparison between RA-ILD and RA-noILD revealed that 2 common IPF risk variants, TOLLIP rs5743890 and IVD rs2034650, were significantly associated with RA-ILD. The TOLLIP rs5743890 minor allele was associated with increased risk of RA-ILD and the IVD rs2034650 minor allele was associated with decreased risk of RA-ILD (ORadj=2.13; 95% CI, 1.13 to 4.10; P=0.02 and ORadj=0.59; 95% CI, 0.38 to 0.89; P=0.01, respectively) and the directionality of these relationships is consistent with what has been observed for IPF.16,17 No association with RA-ILD was detected for the 10 other IPF risk variants (FIG. 29).

Example 8: MUC5B Promoter Variant is Associated with Visually and Quantitatively Detected Preclinical Pulmonary Fibrosis

Better understanding and recognition of early pulmonary fibrosis is critical because medical therapies have been shown to slow progression, not to reverse or even stabilize established fibrosis—therefore, intervention before irreversible fibrosis has become extensive has the potential to improve quality of life and decrease morbidity. While IPF affects approximately 5 million people worldwide, between 1.8 and 14% of the general population ≥50 years of age have radiologic findings of undiagnosed pulmonary fibrosis. Large cohort studies indicate that interstitial lung abnormalities, postulated to represent early pulmonary fibrosis, are associated with increased mortality, and that most of these abnormalities progress over time. Members of families with 2 or more cases of pulmonary fibrosis (FIP, Familial Interstitial Pneumonia) have been identified as an “at-risk” population. In a previous study of FIP relatives, 14% had interstitial lung abnormalities on high resolution computed tomography (HRCT), and 35% had an abnormal transbronchial biopsy indicating interstitial lung disease.

HRCT provides visualization of the lung parenchyma and plays a key role in the diagnosis of the Idiopathic Interstitial Pneumonias (IIPs), including IPF. Currently, visual diagnosis by thoracic radiologists, in conjunction with multidisciplinary clinical conference, is the gold standard for diagnosing IIPs. However, visual assessment is imprecise and hampered by inter-observer variation. Quantitative HRCT (qHRCT) evaluation provides measures of fibrosis extent that, in subjects diagnosed with IPF, correlate with degree of physiologic impairment at baseline, and may be more sensitive to subtle changes in disease status than routinely used physiological metrics. The design and utility of quantitative methods in the context of early forms of fibrotic ILD requires further study. Deep learning methods have been increasingly used in imaging to identify and classify CT patterns, and may be particularly valuable in detection of early lung fibrosis.

This study aims to: (1) examine risk factors, including two common fibrosis-associated genetic variants in MUC5B and TERT, for undiagnosed pulmonary fibrosis (PrePF) in FIP first-degree relatives; and (2) determine the utility of a deep learning, texture-based qHRCT method in the detection of early fibrosis in this cohort.

Materials and Methods FIP Relatives Screening:

As part of a study of FIP conducted at the University of Colorado, National Jewish Health, and Vanderbilt University (COMIRB #15-1147; NJH IRB 1441a; Vanderbilt IRB #020343), non-Hispanic white (NHW) relatives of FIP patients, defined as those in families with two or more cases of pulmonary fibrosis, were contacted for enrollment. First-degree relatives without a known prior diagnosis of pulmonary fibrosis and greater than 40 years of age were offered HRCT scans of the chest and asked to undergo peripheral blood draw. Study subjects younger than 40 years of age or older than 40 years of age who reported on pre-study questionnaires to be personally affected by pulmonary fibrosis were excluded (FIG. 18).

Visual CT Review:

HRCT scans were interpreted by study radiologists and examined for the presence of fibrotic ILD. “PrePF” was defined as the presence of “probable” or “definite” fibrotic ILD on HRCT in FIP relatives who had no known diagnosis of pulmonary fibrosis at the time of study enrollment (FIGS. 18, 19).

Quantitative CT:

Inspiratory HRCT series with slice thickness ≤1·25 mm and spacing ≤20·0 mm were selected for quantitative analysis. This included 212 volumetric series with thin, contiguous sections (slice thickness and spacing both <=1·25 mm) and 191 non-volumetric scans (56 with slice spacing >1·25 mm and <10 mm, 65 with slice spacing of 10 mm and 70 with slice spacing=20 mm). Scans identified as technically inadequate were omitted. In addition, 100 inspiratory volumetric HRCT of never-smoking control subjects from the COPDGene cohort were analyzed (FIG. 20). The lungs were segmented in a semi-automatic fashion using open source software followed by manual editing, if necessary, performed by trained analysts. Examples of the categorization of different parts of CT scans are shown in FIG. 21. Some studies were acquired with contiguous thin axial sections while others used 1 or 2 cm intervals. Also, reconstruction kernel, a parameter that affects image sharpness and noise, was not standardized.

Fibrosis quantification on CT scans was performed using a deep learning technique, with a convolutional neural network (CNN) algorithm trained with image regions of normal and abnormal lung identified by expert radiologists. Training data and an earlier algorithm version were described previously. Here, a more complex CNN architecture was employed that classifies image regions using pixel and texture features extracted by multiple convolutional layers at different scales. Classification categories included normal lung, airways, reticular abnormality, honeycombing and ground glass. An additional category, “not normal”, was also included for lung regions not classified into any of the named categories. Further, pixels in the “not normal” category were split into two subcategories: “not normal” low density and “not normal” high density using the threshold value of −650 Hounsfield Units (HU). Subject level scores were computed as the percentage of total lung volume classified in each category. HRCT fibrosis score was defined as the sum of CNN classification scores for reticular abnormality, honeycombing, ground glass, and “not normal high density” (FIG. 21).

A simpler previously described densitometric analysis of HRCTs was also performed for comparison. Percent high attenuation area (% HAA), the percentage of total lung volume with HRCT pixel intensity greater than −600 HU and less than −250 HU, has been used as a measure of interstitial lung disease on CT.

Statistical Analysis:

Analysis of the effect of specific alleles on PrePF risk was performed using minor allele frequency (MAF) for comparison of variant prevalence in the study groups; statistical significance was determined utilizing either a z-score test for proportions or a mixed effects logistic regression model when controlling for other clinical factors (age, sex, and history of smoking) and family [random effect]) in both dominant and log-additive models.

Distribution of qHRCT fibrosis scores was left skewed as was % HAA, and therefore these values were log transformed prior to analyses. Log of qHRCT fibrosis score (hereafter, “fibrosis score”) and log (% HAA) were compared with visual scores using ANOVA and Tukey's honest significant difference (HSD) test. To determine the ability of qHRCT scores to predict visual diagnosis of PrePF, receiver-operating characteristic (ROC) analysis was performed. Optimal threshold for discriminating visual diagnosis of fibrotic ILD was determined with Youden's method. Five-fold cross-validation was performed to test detection accuracy, sensitivity and specificity, and consistency of optimal threshold. Linear regression was performed to test association between the MUC5B genotype and qHRCT fibrosis score and log (% HAA).

A p-value of <0.05 was considered statistically significant for differences between groups as well as for associations between individual variables and outcomes in linear and logistic regression modeling. Statistical analyses were performed using RStudio (Version 0.99.473).

Results Study Cohort Characteristics

A total of 1,090 FIP relatives were contacted, and 523 eligible subjects were recruited and underwent HRCT screening (FIG. 18). Of the 523 subjects, 26 were excluded due to technical inadequacy of images and one for an equivocal consensus read by study radiologists. The remaining 496 subjects from 263 families were included in the final analyses. The mean age of study subjects was 57 years (95% CI: 56.5-58), 189 (38%) were male, and 148 (29%) were either current or former smokers. The minor allele (T) frequency of the MUC5B promoter polymorphism rs35705950 was 0.22 in this cohort; 45% of the subjects in this cohort had one or two copies of the minor allele (FIG. 22). The minor allele (C) frequency of the TERT variant rs2736100 was 0.47 in the entire cohort; 69% of the subjects in the cohort having one or two copies of the minor allele (FIG. 22).

Prevalence of Preclinical Pulmonary Fibrosis (PrePF) in FIP Relatives

Of the 496 HRCT scans, 401 showed no CT evidence of interstitial lung disease (ILD), and 95 showed evidence of ILD, either fibrotic (27 probable and 50 definite) or non-fibrotic (n=18). Therefore, among these 496 subjects who reported being personally unaffected by pulmonary fibrosis, the PrePF prevalence was 15.5% (n=77) (FIG. 18).

The CT patterns noted in PrePF subjects (FIG. 23) show that possible, probable, or definite UIP pattern was the most commonly considered (n=59, 77% of all PrePF cases). NSIP was considered in 45 subjects (58% of all PrePF cases). The fibrotic changes were most commonly lower-lobe predominant and subpleural in nature, consistent with a UIP pattern (FIG. 23). Non-fibrotic ILD scans, on the other hand, generally had more diffuse, upper-lobe predominant abnormalities.

There were 402 study subjects with HRCT scans that were technically adequate for quantitative assessment. 212 of the scans had both slice thickness and spacing <=1·25 mm (thin, contiguous); of the remaining 191 scans, 56 had slice spacing >1·25 mm and <10 mm, 65 had slice spacing=10 mm, and 70 had slice spacing=20 mm. Volumetric HRCT scans on an additional 100 COPDGene subjects were included as normal controls. Fibrosis score means were significantly different (p<0·0001) across groups defined by visual diagnosis (FIG. 24). Comparison of means showed fibrosis score were significantly different comparing each group (all between-group comparisons p<0·01). Means of log (% HAA) scores were also significantly different across visual scoring groups (p<0·0001), and individual between-group comparisons showed log (% HAA) was significantly different in most comparisons (p<0·0001), except between the “probable” and “definite” visual scores (p=0·35).

ROC analysis showed that fibrosis score discriminates subjects with visual diagnosis of PrePF (FIG. 25B). Average area under the curve (AUC) in five-fold cross validation was 0.85 (range 0.83-0.87) and average accuracy, sensitivity, and specificity in the test partitions were 0.83 (range 0.74-0.86), 0.74 (range 0.56-0.92), and 0.84 (range 0.76-0.89), respectively. Optimal threshold for fibrosis score ranged from 1.40-1.42, corresponding to 4.1% fibrotic area in examined lung. Utilizing a cutoff of 1.40 for fibrosis score on the entire dataset, the sensitivity was 74%, specificity was 82%, and accuracy was 81%; the negative predictive value of this test was 95%, exceeding its positive predictive value (42%) (FIG. 25C).

Compared to the classification achieved with the CNN as described above, ROC analysis of log % HAA had lower mean AUC 0.80 (range 0.79-0.81) and average accuracy, sensitivity, and specificity of 0.67 (range 0.63-0.70), 0.82 (range 0.75-0.91) and 0.64 (range 0.62-0.70) respectively (FIG. 25A). Optimal threshold for log % HAA ranged from 1.49-1.57. Utilizing a cutoff of 1.49 for log % HAA, the sensitivity was 88%, specificity was 55%, and accuracy was 60%; the negative predictive value of this test was 96%, exceeding its positive predictive value (26%).

Risk Factors for PrePF

Subjects with PrePF were older (mean age 65.8 years, 95% CI 63.5-68.1) than those without fibrosis (mean age 55.8, 95% CI 54.9-56.6, p=6.36×10⁻¹³); they were also more likely to have ever smoked (43% versus 27%, p=0·007), and to be male (48% versus 36%, p=0·05). However, there was no difference in breathlessness between the PrePF and subjects without fibrosis (mean score 0.5 versus 0·6, p=0·24, FIG. 26). When fibrosis was defined by quantitative fibrosis score cutoff (1.4), there was a significant difference between groups in terms of mean breathlessness score (0.39 versus 0.78, p=0·003). Quantitative fibrosis score was positively associated with breathlessness score (p=0.001), even after controlling for age (p=1.9×10⁻⁹), male sex (p=0.7), and smoking history (p=0.8).

Screening for autoantibodies in this cohort revealed that there were no differences between PrePF and No Fibrosis subjects in terms of overall seropositivity or individual antibodies' testing in this cohort. For quantitatively defined fibrosis, there was no significant difference between groups in terms of auto-antibody testing, with similar overall seropositivity rates (11% versus 16%, p=0.30).

The MUC5B promoter polymorphism rs35705950 was associated with the visual diagnosis of PrePF (present in 40% of those without fibrosis versus 53% with PrePF; MAF 0.29 versus 0.21, respectively, p=0.03, FIG. 22). After age 60, there was a statistically significant difference in the proportion of subjects with visually diagnosed PrePF when the cohort was stratified by MUC5B genotype (23.8% versus 39.8% prevalence, p=0.02) (FIG. 27).

MUC5B variant carriers, regardless of their visual CT diagnosis, had significantly higher qHRCT fibrosis scores (1.3 [95% CI 1.2-1.5] versus 1.1 [95% CI 1.0-1.2], p=0.02). The association between MUC5B genotype and fibrosis score was significant even when controlling for age and male sex in linear regression (p=0.03, FIG. 28). Age was significantly associated with fibrosis score (p=2.17×10⁻⁹), but male sex (p=0.63) and smoking (p=0.94) were not. To determine whether individual textural components were driving the association of the composite fibrosis score with genotype, each score component was tested individually for association with the MUC5B variant, controlling for age and sex. Quantitative scores for reticulation, honeycombing, and ground glass were significantly associated with the MUC5B variant (p=0.02, p=0.02, p=0·04, respectively), while “not normal high density” was not (p=0·18). The simpler quantitative scoring method, log % HAA, was not significantly different in MUC5B variant carriers (p=0.4).

In contrast to the MUC5B variant, the common IPF-associated TERT polymorphism (rs2736100) was not significantly associated with PrePF assessed either qualitatively (MAF 0.47 in PrePF versus 0.46 in unaffected, p=0.77) or quantitatively (MAF 0.50 fibrotic versus 0.47 not fibrotic, p=0.40).

When these factors were examined individually for their contributions to risk of PrePF in our study cohort, we used a mixed effects logistic regression model to test the independent effects of age sex, smoking, and MUC5B or TERT genotypes while controlling for family. Age and the MUC5B genotype remained statistically significantly associated with PrePF (OR 1.15, 95% CI 1.09-1.22, p=7.34×10⁻⁷ and OR 2.18, 95% CI 1.00-4.73, p=0.05, respectively) (FIG. 22). The common TERT polymorphism (rs2736100) associated with fibrotic idiopathic interstitial pneumonia (29) was not significantly associated with PrePF (MAF was 0.45 in PrePF versus 0.45 in unaffected, p=0.88) or in a log-additive model controlling for age, sex, and smoking history (p=0.57).

Given the presence of non-fibrotic ILD (n=18, FIG. 18) in the “No Fibrosis” cohort, secondary analyses were performed that (1) excluded non-fibrotic ILDs and (2) compared all ILD (inclusive of non-fibrotic ILD) to those without any ILD. When non-fibrotic ILDs were excluded from analyses, PrePF subjects were older (p=4.7×10⁻¹³), more commonly male (p=0.04), more often had a smoking history (p=0.003), and had a higher prevalence of the MUC5B promoter variant (MAF 0.29 versus 0.20, p=0.02). However, when controlling for family relatedness and the other risk factors in a mixed effects logistic regression, only age and the MUC5B promoter variant were significantly associated with PrePF with odds ratios 1.15 (95% CI 1.09-1.22, p=9.5×10⁻⁷) and 2.16 (95% CI 1.00-4.75, p=0.05), respectively. Another secondary analysis of the data was performed in which all subjects with CT findings of ILD (fibrotic or non-fibrotic) were compared to those without any evidence of ILD. Those with CT evidence of ILD were older (mean age 64.3 years, 95% CI 62.2-66.3) compared to those without any evidence of ILD (mean age 55.7 years, 95% CI 54.8-56.6, p=4.1×10⁻¹²), more likely to be male (p=0.01), more likely to have smoked (p=0.0003), and more likely to carry the MUC5B promoter variant (MAF 0.21 versus 0.30, p=0.006). When controlling for family relatedness in a mixed effects logistic regression model, age (OR 1.10, 95% CI 1.07-1.14, p=1.21×10⁻⁹), smoking history (OR 1.72, 95% CI 1.00-2.99, p=0.04), and the MUC5B promoter variant (OR 1.73, 95% CI 1.08-2.76, p=0·02) were significantly associated with risk of ILD.

OTHER EMBODIMENTS

It is to be understood that while the disclosure has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the disclosure, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. 

What is claimed is:
 1. A method of treating a fibrotic lung disease in a subject comprising administering to the subject an effective amount of a therapeutic agent, wherein the subject is asymptomatic and wherein the subject is at risk of developing the fibrotic lung disease.
 2. The method of claim 1, wherein the subject presents radiographic Usual Interstitial Pneumonia (UIP).
 3. The method of claim 1 or 2, wherein the subject has fibrotic interstitial lung disease (FILD).
 4. The method of claim 1 or 2, wherein the subject has an interstitial lung disease (ILD).
 5. The method of claim 4, where in the subject has rheumatoid arthritis-associated interstitial lung disease (RA-ILD).
 6. The method of any one of claims 1-3, wherein the subject has a blood relative with familial interstitial pneumonia (FIP).
 7. The method of claim 6, wherein the blood relative is a sibling.
 8. The method of any one of claims 1-7, wherein the subject has a mutation in a sequence encoding Mucin 5B (MUC5B), Telomerase RNA Component (TERC), Family with sequence similarity 13 member A (FAM13A), Telomerase Reverse Transcriptase (TERT), Desmoplakin (DSP), Zinc-alpha 2-Glycoprotein 1 (AZGP1), Oligonucleotide/oligosaccharide-binding Fold Containing 1 (OBFC1), ATPase Phospholipid Transporting 11A (ATP11A), Isovaleryl-CoA dehydrogenase (IVD)/Dispatched RND Transporter Family Member 2 (DISP2), Dipeptidyl Peptidase 9 (DPP9), Sialic Acid Binding Ig-Like Lectin 14 (SIGLEC14), Adrenomedullin 2 (ADM2), Tetraspanin 5 (TSPAN5), Calcium/Calmodulin-Dependent Protein Kinase Kinase 1 (CAMKK1), zinc figner with KRAB and SCAN domains 1 (ZKSCAN1), isovaleryl-CoA dehydrogenase (IVD), ATPase phospholipid transporting 11A (AK025511) or Matrix Metalloprotease-7 (MMP-7).
 9. The method of claim 8, wherein the mutation comprises a polymorphism in a sequence encoding a MUC5B promoter.
 10. The method of claim 9, wherein the polymorphism is rs35705950.
 11. The method of claim 8, wherein the mutation comprises a polymorphism in a sequence encoding a TERC 3′ untranslated region (UTR).
 12. The method of claim 11, wherein the polymorphism is rs2293607.
 13. The method of claim 8, wherein the mutation comprises a polymorphism in a sequence encoding intronic FAM13A.
 14. The method of claim 13, wherein the polymorphism is rs2609260.
 15. The method of claim 8, wherein the mutation comprises a polymorphism in a sequence encoding intronic TERT.
 16. The method of claim 15, wherein the polymorphism is rs4449583.
 17. The method of claim 8, wherein the mutation comprises a polymorphism in a sequence encoding intronic DSP.
 18. The method of claim 17, wherein the polymorphism is rs2076295.
 19. The method of claim 8, wherein the mutation comprises a polymorphism in a sequence encoding intronic ZKSCAN1.
 20. The method of claim 19, wherein the polymorphism is rs6963345.
 21. The method of claim 8, wherein the mutation comprises a polymorphism in a sequence encoding intronic OBFC1.
 22. The method of claim 21, wherein the polymorphism is rs2488000.
 23. The method of claim 8, wherein the mutation comprises a polymorphism in a sequence encoding an AK025511 3′ UTR.
 24. The method of claim 23, wherein the polymorphism is rs1278769.
 25. The method of claim 8, wherein the mutation comprises a polymorphism in a sequence encoding IVD.
 26. The method of claim 25, wherein the polymorphism is rs35700143.
 27. The method of claim 8, wherein the mutation comprises a polymorphism in a sequence encoding intronic DPP9.
 28. The method of claim 27, wherein the polymorphism is rs12610495.
 29. The method of any one of claims 1-28, wherein the fibrotic lung disease is pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), an interstitial lung abnormality (ILA), an asymptomatic ILA, interstitial lung disease (ILD) or rheumatoid arthritis-associated interstitial lung disease (RA-ILD).
 30. The method of any one of claims 1-28, wherein the fibrotic lung disease is ILD or RA-ILD.
 31. The method of any one of claims 1-28, wherein the fibrotic lung disease is pulmonary fibrosis or IPF.
 32. The method of any one of claims 1-28, wherein the fibrotic lung disease is IPF.
 33. The method of any one of claims 1-32, wherein the therapeutic agent comprises a N-acetylcysteine, pirfenidone, and nintedanib.
 34. The method of any one of claims 1-32, wherein the therapeutic agent comprises pirfenidone.
 35. The method of claim 34, wherein the effective dosage is about 2400 mg/day.
 36. The method of claim 35, wherein the effective dosage is administered orally as a capsule or a tablet.
 37. The method of claim 35 or 36, wherein the effective dosage is administered three times per day.
 38. The method of any one of claims 35-37, wherein the effective dosage is administered according to an escalating dosage regimen.
 39. The method of claim 39, wherein the escalating dosage regimen comprises (a) administering to the subject about 800 mg of pirfenidone per day for a first week; (b) administering to the subject about 1600 mg of pirfenidone per day for a second week; and (c) administering to the subject about 2400 mg of pirfenidone per day for the remainder of the treatment.
 40. The method of claim 38, wherein the escalating dosage regimen comprises (a) administering to the subject a capsule or tablet comprising about 250 mg of pirfenidone three times a day for a first week; (b) administering to the subject two capsules or tablets comprising about 250 mg of pirfenidone three times a day for a second week; and (c) administering to the subject three capsules or tablets comprising about 250 mg of pirfenidone three times a day for the remainder of the treatment.
 41. The method of claim 40, wherein the capsule or tablet comprises 267 mg of pirfenidone.
 42. The method of any one of claims 1-32, wherein the therapeutic agent comprises nintedanib.
 43. The method of claim 42, wherein the effective dosage is administered orally as a capsule or a tablet.
 44. The method of claim 43, wherein the effective dosage is about 300 mg/day.
 45. The method of claim 42 or 43, wherein the effective dosage is about 150 mg administered twice per day, wherein the daily doses are administered about 12 hours apart from one another.
 46. The method of claim 43, wherein the effective dosage is about 200 mg/day.
 47. The method of claim 43 or 46, wherein the effective dosage is about 100 mg administered twice per day, wherein the daily doses are administered about 12 hours apart from one another.
 48. The method of any one of claims 42-47, wherein the effective dosage is administered according to a modified or interrupted dosage regimen.
 49. The method of claim 48, wherein the modified or interrupted dosage regimen comprises (a) administering to the subject about 300 mg of nintedanib per day until the subject presents an elevated level of liver enzymes compared to a control level of liver enzymes; (b) administering to the subject about 200 mg of nintedanib per day until the subject presents the control level of liver enzymes; and (c) administering to the subject about 300 mg of nintedanib per day for the remainder of the treatment; wherein the control level of liver enzymes is a level detected in the subject prior to an initiation of the treatment.
 50. The method of claim 48, wherein the modified or interrupted regimen comprises (a) administering to the subject a capsule or tablet comprising about 150 mg of nintedanib twice per day until the subject presents an elevated level of liver enzymes compared to a control level of liver enzymes; (b) administering to the subject two capsules or tablets comprising about 100 mg twice per day until the subject presents an elevated level of liver enzymes compared to a control level of liver enzymes; and (c) administering to the subject a capsule or tablet comprising about 150 mg of nintedanib twice per day for the remainder of the treatment; wherein the control level of liver enzymes is a level detected in the subject prior to an initiation of the treatment.
 51. The method of any one of claims 1-50, wherein the therapeutic agent prevents the onset or development of a sign or symptom of the fibrotic lung disease.
 52. The method of any one of claims 1-50, wherein the therapeutic agent delays the onset or development of a sign or symptom of the fibrotic lung disease when compared to the expected onset of the a sign or symptom in the absence of treatment with the therapeutic agent.
 53. The method of any one of claims 1-50, wherein the therapeutic agent reduces the severity of a sign or symptom of the fibrotic lung disease when compared to the expected severity of the a sign or symptom in the absence of treatment with the therapeutic agent.
 54. The method of any one of claims 51-53, wherein at least one sign of the fibrotic lung disease is detectable before the subject presents a symptom of the fibrotic lung disease.
 55. The method of claim 54, wherein the at least one sign comprises gradual or unintended weight loss, clubbing of the fingers or toes, rapid and shallow breathing, fibrotic lesions in one or both lungs detectable by radiography, or a cough.
 56. The method of claim 54, wherein the symptom comprises shortness of breath during exercise, shortness of breath at rest, a dry and hacking cough, repeated bouts of coughing, and uncontrollable bouts of coughing.
 57. The method of any one of claims 1-56, wherein the method prevents the onset of a secondary condition associated with a severe form of the fibrotic lung disease.
 58. The method of claim 57, wherein secondary condition comprises a collapsed lung, an infected lung, a blood clot in a lung, lung cancer, respiratory failure, pulmonary hypertension, heart failure or death.
 59. A method of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease, comprising administering to a non-human subject a dose of a composition that modifies transcription or translation of a sequence encoding Mucin 5B (MUC5B), Telomerase RNA Component (TERC), Family with sequence similarity 13 member A (FAM13A), Telomerase Reverse Transcriptase (TERT), Desmoplakin (DSP), Zinc-alpha 2-Glycoprotein 1 (AZGP1), Oligonucleotide/oligosaccharide-binding Fold Containing 1 (OBFC1), ATPase Phospholipid Transporting 11A (ATP11A), Isovaleryl-CoA dehydrogenase (IVD)/Dispatched RND Transporter Family Member 2 (DISP2), Dipeptidyl Peptidase 9 (DPP9), Sialic Acid Binding Ig-Like Lectin 14 (SIGLEC14), Adrenomedullin 2 (ADM2), Tetraspanin 5 (TSPAN5), Calcium/Calmodulin-Dependent Protein Kinase Kinase 1 (CAMKK1) or Matrix Metalloprotease-7 (MMP-7), wherein the dose of the composition is tolerable to the non-human subject and wherein the dose of the composition is therapeutically effective.
 60. A method of identifying a therapeutic agent or target thereof for the treatment of a fibrotic lung disease, comprising administering to a non-human subject a composition that modifies an activity of a product of a sequence encoding MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7, wherein the dose of the composition is tolerable to the non-human subject and wherein the dose of the composition is therapeutically effective.
 61. The method of claim 59, wherein the composition that modifies transcription or translation decreases or inhibits transcription or translation.
 62. The method of claim 61, wherein the composition decreases or inhibits transcription or translation of a sequence encoding a gene selected from the group consisting of Leukotriene A4 Hydrolase (LTA4H), Surfactant Protein B (SFTPB), Breast Cancer Anti-Estrogen Resistance 3 (BCAR3), C-X-C motif Chemokine Ligand 13 (CXCL13), EPH Receptor A2 (EPHA2), Serum Amyloid A1 (SAA1), Phospholipase A2 Group IIA (PLA2G2A), Insulin-Like Growth Factor Binding Protein 3 (IGFBP3), C-C Motif Chemokine Ligand 28 (CCL28), S100 Calcium Binding Protein A12 (S100A12), Thromboxane A Synthase 1 (TBXAS1), Leukocyte Cell Derived Chemotaxin 1 (LECT1), Complement C3 (C3), Gastrin Releasing Peptide (GRP), C-Reactive Protein (CRP), Vitrin (VIT), Insulin-Like Growth Factor Binding Protein 1 (IGFBP1), Family with Sequence Similarity 173 Member A (FAM173A), Natriuretic Peptide A (NPPA), Secreted Frizzled Related Protein 1 (SFRP1), Ezrin (EZR), Inter-Alpha-Trypsin Inhibitor Heavy Chain Family Member 5 (ITIH5), Pleckstrin and Sec7 Domain Containing 2 (PSD2), Galectin 3 Binding Protein (LGALS3BP), Catenin Beta 1 (CTNNB1), Chromodomain Y Like 2 (CDYL2), Matrix Metallopeptidase 7 (MMPI), Apolipoprotein B (APOB), Proline and Arginine Rich End Leucine Rich Repeat Protein (PRELP), Eukaryotic Translation Initiation Factor 1A, X-linked (EIF1AX), Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF), TNF Receptor Superfamily Member 13C (TNFRSF13C), Deformed Epidermal Autoregulatory Factor 1 transcription factor (DEAF1), Tumor Protein Translationally-Controlled 1 (TPT1), Unc-5 Netrin Receptor B (UNCSB), Phosphatidylethanolamine Binding Protein 1 (PEBP1), Syntaxin 8 (STX8), Polymeric Immunoglobulin Receptor (PIGR), Adenine Phosphoribosyltransferase (APRT), Matrix Metallopeptidase 3 (MMP3), Galectin 7 (LGALS7), Bruton Tyrosine Kinase (BTK), NSFL1 Cofactor (NSFL1C), FER Tyrosine Kinase (FER), Regenerating Family Member 1 Beta (REG1B), SMAD Family Member 2 (SMAD2), Interleukin 1 Receptor Like 1 (IL1RL1), C-C Motif Chemokine Ligand 18 (CCL18), Acid Phosphatase 2 Lysosomal (ACP2), Eukaryotic Translation Initiation Factor 4E Family Member 2 (EIF4E2), Neurexin 3 (NRXN3), IGF Like Family Member 1 (IGFL1), NME/NM23 Nucleoside Diphosphate Kinase 1 (NME1), Potassium Voltage-Gated Channel Isk-Related Family Member 1-Like (KCNE1L) or Neurexophilin 2 (NXPH2).
 63. The method of claim 59, wherein the composition that modifies transcription or translation increases or activates transcription or translation.
 64. The method of claim 63, wherein the composition increases or activates transcription or translation of a sequence encoding a gene selected from the group consisting of Surfactant Protein D (SFTPD), Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), Histone Cluster 1 H1 Family Member C (HIST1H1C), YTH Domain Containing 1 (YTHDC1), Plexin A1 (PLXNA1), Serine Peptidase Inhibitor Kazal Type 6 (SPINK6), LDL Receptor Related Protein Associated Protein 1 (LRPAP1), Secretoglobin Family 3A Member 1 (SCGB3A1), H2A Histone Family Member Z (H2AFZ) or Chromosome 1 Open Reading Frame 162 (C1 orf162).
 65. The method of claim 60, wherein the composition that modifies an activity decreases or inhibits the activity.
 66. The method of claim 65, wherein the composition decreases or inhibits the activity of a sequence encoding a gene selected from Leukotriene A4 Hydrolase (LTA4H), Surfactant Protein B (SFTPB), Breast Cancer Anti-Estrogen Resistance 3 (BCAR3), C-X-C motif Chemokine Ligand 13 (CXCL13), EPH Receptor A2 (EPHA2), Serum Amyloid A1 (SAA1), Phospholipase A2 Group IIA (PLA2G2A), Insulin-Like Growth Factor Binding Protein 3 (IGFBP3), C-C Motif Chemokine Ligand 28 (CCL28), 5100 Calcium Binding Protein A12 (S100A12), Thromboxane A Synthase 1 (TBXAS1), Leukocyte Cell Derived Chemotaxin 1 (LECT1), Complement C3 (C3), Gastrin Releasing Peptide (GRP), C-Reactive Protein (CRP), Vitrin (VIT), Insulin-Like Growth Factor Binding Protein 1 (IGFBP1), Family with Sequence Similarity 173 Member A (FAM173A), Natriuretic Peptide A (NPPA), Secreted Frizzled Related Protein 1 (SFRP1), Ezrin (EZR), Inter-Alpha-Trypsin Inhibitor Heavy Chain Family Member 5 (ITIH5), Pleckstrin and Sec7 Domain Containing 2 (PSD2), Galectin 3 Binding Protein (LGALS3BP), Catenin Beta 1 (CTNNB1), Chromodomain Y Like 2 (CDYL2), Matrix Metallopeptidase 7 (MMPI), Apolipoprotein B (APOB), Proline and Arginine Rich End Leucine Rich Repeat Protein (PRELP), Eukaryotic Translation Initiation Factor 1A, X-linked (EIF1AX), Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF), TNF Receptor Superfamily Member 13C (TNFRSF13C), Deformed Epidermal Autoregulatory Factor 1 transcription factor (DEAF1), Tumor Protein Translationally-Controlled 1 (TPT1), Unc-5 Netrin Receptor B (UNCSB), Phosphatidylethanolamine Binding Protein 1 (PEBP1), Syntaxin 8 (STX8), Polymeric Immunoglobulin Receptor (PIGR), Adenine Phosphoribosyltransferase (APRT), Matrix Metallopeptidase 3 (MMP3), Galectin 7 (LGALS7), Bruton Tyrosine Kinase (BTK), NSFL1 Cofactor (NSFL1C), FER Tyrosine Kinase (FER), Regenerating Family Member 1 Beta (REG1B), SMAD Family Member 2 (SMAD2), Interleukin 1 Receptor Like 1 (IL1RL1), C-C Motif Chemokine Ligand 18 (CCL18), Acid Phosphatase 2 Lysosomal (ACP2), Eukaryotic Translation Initiation Factor 4E Family Member 2 (EIF4E2), Neurexin 3 (NRXN3), IGF Like Family Member 1 (IGFL1), NME/NM23 Nucleoside Diphosphate Kinase 1 (NME1), Potassium Voltage-Gated Channel Isk-Related Family Member 1-Like (KCNE1L) or Neurexophilin 2 (NXPH2).
 67. The method of claim 60, wherein the composition that modifies an activity increases or activates the activity.
 68. The method of claim 67, wherein the composition increases or activates the activity of a sequence encoding Surfactant Protein D (SFTPD), Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), Histone Cluster 1 H1 Family Member C (HIST1H1C), YTH Domain Containing 1 (YTHDC1), Plexin A1 (PLXNA1), Serine Peptidase Inhibitor Kazal Type 6 (SPINK6), LDL Receptor Related Protein Associated Protein 1 (LRPAP1), Secretoglobin Family 3A Member 1 (SCGB3A1), H2A Histone Family Member Z (H2AFZ) or Chromosome 1 Open Reading Frame 162 (Clorf162).
 69. The method of any one of claims 60-68, wherein the non-human subject is a mammal.
 70. The method of any one of claims 60-69, wherein the mammal is genetically-modified.
 71. The method of claim 70, wherein the genetically-modified mammal is a model organism for the fibrotic lung disease.
 72. The method of any one of claims 60-71, wherein the fibrotic lung disease is pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), an interstitial lung abnormality (ILA), or an asymptomatic ILA.
 73. The method of any one of claims 60-71, wherein the fibrotic lung disease is pulmonary fibrosis or IPF.
 74. The method of any one of claims 60-71, wherein the fibrotic lung disease is IPF.
 75. The method of any one of claims 60-74, wherein the non-human subject carries a mutation in a sequence encoding MUC5B.
 76. The method of claim 75, wherein the mutation comprises a polymorphism in a sequence encoding a MUC5B promoter.
 77. The method of claim 76, wherein the polymorphism is rs35705950.
 78. The method of any one of claims 60-77, wherein the non-human subject carries a mutation in a sequence encoding TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7.
 79. The method of any one of claims 60-78, wherein the composition prevents the onset or development of a sign or symptom of the fibrotic lung disease.
 80. The method of any one of claims 60-78, wherein the composition delays the onset or development of a sign or symptom of the fibrotic lung disease when compared to the expected onset of the a sign or symptom in the absence of treatment with the composition.
 81. The method of claim 80, wherein the composition delays the onset or development of a sign or symptom of the fibrotic lung disease when compared to the expected onset of the sign or symptom when treated using a standard therapeutic intervention.
 82. The method of any one of claims 60-78, wherein the composition reduces the severity of a sign or symptom of the fibrotic lung disease when compared to the expected severity of the sign or symptom in the absence of treatment with the composition.
 83. The method of claim 24, wherein the composition reduces the severity of a sign or symptom of the fibrotic lung disease when compared to the expected severity of the sign or symptom when treated using a standard therapeutic intervention.
 84. The method of claim 81 or 83, wherein the standard therapeutic intervention comprises a N-acetylcysteine, pirfenidone, and nintedanib.
 85. The method of claim 81 or 83, wherein the standard therapeutic intervention comprises pirfenidone.
 86. The method of claim 85, wherein an effective dosage of pirfenidone is about 2400 mg/day.
 87. The method of claim 86, wherein the effective dosage is administered orally as a capsule or a tablet.
 88. The method of claim 86 or 87, wherein the effective dosage is administered three times per day.
 89. The method of any one of claims 85-88, wherein the effective dosage is administered according to an escalating dosage regimen.
 90. The method of claim 85, 86 or 89, wherein the escalating dosage regimen comprises (a) administering to the non-human subject about 800 mg of pirfenidone per day for a first week; (b) administering to the non-human subject about 1600 mg of pirfenidone per day for a second week; and (c) administering to the non-human subject about 2400 mg of pirfenidone per day for the remainder of the treatment.
 91. The method of claim 85, 86 or 89, wherein the escalating dosage regimen comprises (a) administering to the non-human subject a capsule or tablet comprising about 250 mg of pirfenidone three times a day for a first week; (b) administering to the non-human subject two capsules or tablets comprising about 250 mg of pirfenidone three times a day for a second week; and (c) administering to the non-human subject three capsules or tablets comprising about 250 mg of pirfenidone three times a day for the remainder of the treatment.
 92. The method of claim 91, wherein the capsule or tablet comprises 267 mg of pirfenidone.
 93. The method of claim 81 or 83, wherein the standard therapeutic intervention comprises nintedanib.
 94. The method of claim 93, wherein an effective dosage of nintedanib is administered orally as a capsule or a tablet.
 95. The method of claim 94, wherein the effective dosage is about 300 mg/day.
 96. The method of claim 94 or 95, wherein the effective dosage is about 150 mg administered twice per day, wherein the daily doses are administered about 12 hours apart from one another.
 97. The method of claim 94, wherein the effective dosage is about 200 mg/day.
 98. The method of claim 94 or 95, wherein the effective dosage is about 100 mg administered twice per day, wherein the daily doses are administered about 12 hours apart from one another.
 99. The method of any one of claims 79-98, wherein the non-human subject presents at least one sign of the fibrotic lung disease.
 100. The method of claim 99, wherein the at least one sign comprises gradual or unintended weight loss, clubbing of the fingers or toes, rapid and shallow breathing, fibrotic lesions in one or both lungs detectable by radiography, or a cough.
 101. The method of any one of claims 60-100, wherein the compound prevents the onset of a secondary condition associated with a severe form of the fibrotic lung disease.
 102. The method of claim 100, wherein the compound prevents the onset for at 1 year, 2 years, 3 years, 4 years, 5 years or any whole or fractional number of years in between.
 103. The method of claim 101 or 102, wherein secondary condition comprises a collapsed lung, an infected lung, a blood clot in a lung, lung cancer, respiratory failure, pulmonary hypertension, heart failure or death.
 104. A composition for the treatment of a fibrotic lung disease identified by the method of any one of claims 60-103.
 105. A method of treating a fibrotic lung disease in a human subject comprising administering to the subject the composition of claim 104, wherein the subject is asymptomatic and wherein the subject is at risk of developing the fibrotic lung disease.
 106. The method of claim 105, wherein the human subject presents radiographic Usual Interstitial Pneumonia (UIP).
 107. The method of claim 105 or 106, wherein the human subject has fibrotic interstitial lung disease (FILD).
 108. The method of any one of claims 105-107, wherein the human subject has a blood relative with familial interstitial pneumonia (FIP).
 109. The method of claim 108, wherein the blood relative is a sibling.
 110. The method of any one of claims 105-109, wherein the human subject has a mutation in a sequence encoding MUC5B, TERC, FAM13A, TERT, DSP, AZGP1, OBFC1, ATP11A, IVD/DISP2, DPP9, SIGLEC14, ADM2, TSPAN5, CAMKK1 or MMP-7.
 111. The method of claim 110, wherein the mutation comprises a polymorphism in a sequence encoding a MUC5B promoter.
 112. The method of claim 111, wherein the polymorphism is rs35705950.
 113. The method of any one of claims 105-112, wherein the fibrotic lung disease is pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), an interstitial lung abnormality (ILA), or an asymptomatic ILA.
 114. The method of any one of claims 105-112, wherein the fibrotic lung disease is pulmonary fibrosis or IPF.
 115. The method of any one of claims 105-112, wherein the fibrotic lung disease is IPF.
 116. The method of any one of claims 105-112, wherein the method prevents the onset of a secondary condition associated with a severe form of the fibrotic lung disease.
 117. The method of claim 116, wherein secondary condition comprises a collapsed lung, an infected lung, a blood clot in a lung, lung cancer, respiratory failure, pulmonary hypertension, heart failure or death. 